Acta Biotechnologica: Volume 3, Number 4 [Reprint 2021 ed.] 9783112542903, 9783112542897

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Volume 3 • 1983 • Number 4

Journal of microbial, biochemical and bioanalogous technology

Akademie-Veriag Berlin ISSN 0138-4988 Acta Biotechnol., Berlin 3 (1983) A, 297-382 EVP 3 0 , - M

Instructions to Authors

1. Only original papers that have not previously been published will be accepted. Manuscripts may be submitted in English, German or Russian but always in duplicate. The name of the institute (inclusive full address) from which the manuscript originates should be stated below the author's name(s). The authors are responsible for the contents of their contributions. 2. Original papers should not consist of more than 20 typewritten pages (2 lined-spacing), including references, tables and figures; short original communications may have a maximum of 6 typewritten pages. 3. Each paper should be preceded by a summary in english. 4. Latin names of species as well as passages to be printed in italics for greater emphasis should be signed with weved line. Please use only units and symbols of the Si-system. 5. Tables may be used to effect a reduction of the text or to make assimilation easier. They should be numbered consecutively throughout and supplied with a brief heading. They shouldn't be written among the text but on separate sheets. 6. Number and size of illustrations should be limited to the essential minimum, they should be numbered consecutively and brought on separate sheets. Line drawings including graphs and diagrams should be drawn in black ink. Half-tone illustrations should be supplied as white glossy prints. Figure legends are to be typed in sequence on a separate sheet. Authors name is to be written on to the rear of each sheet. 7. References, listed at the end of the contribution, should contain only works cited in the text. They should be numbered in the order in which they are at first mentioned in the text. Please give surnames and initials of all authors, name of journal abbreviated according to "Chemical Abstracts — List of Periodicals", volume number, year of publication, issue number or month, first page number; books are to be cited with full title, edition, volume number, page number, place of publication, publisher and year of publication. 8. Notes on the text may be given as footnotes on the relevant page. 9. 50 reprints are free of charge. Additional reprints may be ordered. 10. The author receives 2 galley proofs for correction. They are sent back to the redaction (Dr. Dimter, DDR-7060 Leipzig, Permoserstr. 15) as soon as possible.

Ada Blitecbnmiica Journal of microbial, biochemical and bioanalogous technology

Volume 3 • 1983

AKADEMIE-VERLAG

Edited at Institute of Technical Chemistry of the Academy of Sciences of the G.D.R.; Leipzig and Institute of Technical Microbiology; Berlin by M. Ringpfeil, Leipzig and G. Vetterlein, Berlin

Editorial board: M. E. Beker, Riga H . W. Blanch, Berkeley S. Fukui, Kyoto H. G. Gyllenberg, Helsinki G. Hamer, Zurich J . Hollo, Budapest M. V. Iwanow, Pushchino F. J u n g , Berlin H . W. D. Katinger, Vienna K . A. Kalunjanz, Moscow J . M. Lebeault, Compiegne D. Meyer, Leipzig

P . Moschinski, Lodz A. Moser, Graz M. D. Nicu, Bukarest Chr. Panajotov, Sofia L. D. Phai, Hanoi H. Sahm, Jülich W. Scheler, Berlin R . Schulze, K o t h e n B. Sikyta, Prague G. K . Skrjabin, Moscow M. A. Urrutia, Habana J . E. Zajic, El Paso

Redaction:

L. Dimter, Leipzig

•

B E R L I N

2

Contents of Volume 3

Contents of Volume 3 (1983) Number 1 . 1 9 8 3 K. H. STEINKRAUS: Traditional Food Fermentations as Industrial Resources (Eng.) . . . . R. H Ä U S E L , R. H E D L I C H and M. J E C H O R E K : Growth Characteristics of a Thermotolerant Strain of Lodderomyces elongisporus grown on Sucrose (Eng.)

J . D . SCHNEIDER, K . H . HOFMANN

and

W. BABEL:

Methylglyoxal—a toxic Fermentation Product (Ger.) . . .

3 13 21

M. P. L A I T E : Changes of the Genetic Patterns and Microevolutional Processes under the Conditions of an intensive Lysine Production at some Strains of Brevibacterium (Russ.) . . . .

27

and P. W E L O N O W : Effect of the Chemical Insecticide Fekamatribufon EC 50 on Acricultural Pesticides obtained from Bacillus thuringiensis (Ger.) . .

33

A. T Ä U F E L and H . R U T T L O F F : Production and Characterization of Proteases of Thermoactinomyces vulgaris (Ger.)

39

and K . M A L I K : Overproduction of Aminoacids and the Activity of Methanol Dehydrogenase of Protaminobacter thiaminophagus Mutants (Eng.)

49

A. OPOKU and Amylase (Eng.)

Saccharification of Cassava (Manihot esculenta) with Millet '

53

E. W . J W A N N Y and M . M . RASHAD : Sugar Compositions of Extra- and Intracellular Polysaccharides produced by Candida lipolytica (4124) grown on n-Hexadecane (Eng.)

59

V. W. OGUNDERO : Factors affecting Growth and Cellulose Hydrolysis by the Thermotolerant Aspergillus nidulans from Composts (Eng.)

65

S. KONÉ and U. BEHRENS: The Maintenance Metabolism and the "maximum" Yield Value (Ger.)

73

S.

76

S . ILIEWA,

I W . STOJANOW

A . LEUCHTENBERGER,

Z . L I B U D Z I S Z , H . OBERMAN

KONÉ

and U.

N. URAIH:

BEHRENS:

Meaning of the True Maximum Specific Growth Rate (Ger.) . .

A . SCHMIDT, D . KUCHENBECKER and H . DÄBERITZ : P r e l i m i n a r y R e s u l t s in t h e F l u o r o m e t r i c

Investigation of Fermentation Processes (Eng.)

,

79

B. H E I N R I T Z , P. STOLL and F. GLOMBITZA: Heat Flow Measurements in Aerobic Microbial Growth Processes with a Nonisothermal Calorimeter operating directly (Eng.) . . . .

83

and R . D . R E C K N A G E L : A new Biometrical Screening Method as applied to Nutrient Optimization in Fermentation (Eng.)

89

H . BOCKER

H . - P . SCHMAUDER, D . GRÖBER and W . BADE: Construction of B i o r e a c t o r s suited for the

Cultivation of Plant Cells (Ger.)

93

Number 2 . 1 9 8 3 E. G A R A Y , E. A L C A I D E and Physiology and Industrial Applications (Eng.)

J.P.MARTÍNEZ,

E.HERNÁNDEZ:

The Genus

Thiobacillus:

99

E. S T I C H E L and T H . B L E Y : Use of Biorhythmic Processes for Increasing the Efficiency of Carbon-Compound Conversion by Microorganisms (Eng.) 125 B . HEINRITZ, G. ROGGE,

H. L U T H E R , A. CLAUS, W. Yeast Cells (Ger.)

BÜTTNER

and E.

SCHUSTER:

Separation of mechanically disrupted

133

M. BULLMANN, G . - J . U H L E N H U T and G . S C H I N D L E R : Kinetics of Mixed Substrates at Citric Acid Accumulation by Saccharomycopsis lipolytica EH 59 (Ger.) . 143 U . ISKE,

CH. G W E N N E R ,

and I. M. C H I R K O V : Electrophysical Methods of Monitoring Biosynthesis (Eng.) 1 5 5 E. DANIELOVÁ, V . FARK AS and S. B A U E R : Enzymatic Hydrolysis and Ethanol Fermentation of Lignocellulosic Materials (Ger.) 163 N. V . SEDYKH

A. SCHMIDT: The Technological Possibilities for Measuring of Physiological Population State Features in Fermentation Practice (Eng.) 171

Contents of Volume 3

3

LB dinh PHAI: Biochemical and Microbiological Aspects of the " N u o c - m a m " I n d u s t r y in Vietnam (Ger.) 177 and H . P . K L E B E R : Regulation of t h e extracellular Lipase from an Alkan Utilizing Strain of Acinetobacter (Ger.) 185 D . HAJERBURG

dumber 3.1983 A. MOSER: Formal Macroapproach to Bioprocessing-Modeling with Analogies (Eng.) . . . .

195

S . K . SEN, M . CHATTERJEE, S . P . CHATTERJEE, A . K . B A N E R J E E : A c t i v i t y of D i a m i n o p i m e l i c

Acid Decarboxylase in Bacteria Producing Lysine (Eng.)

217

E. E M A N U I L O V A , A. K A I M A K T C H I E V : Production of Neutral and Alkaline Proteases in Batch and Chemostat Cultures by Bacillus mesentericus 76 (Eng.) 221 M. RINGPFEIL: SCP-Production on the Basis of Hydrocarbons (Ger.)

227

N . B . GRADOVA, E . M . DIKANSKAYA, S . N . ROBYCHEVA, G . S . RODIONOVA, M . B . BITTEYEVA,

: Characteristics of Yeast Strains for t h e Production of SCP on the Basis of Petroleum Hydrocarbons. Specific Growth Properties and Biosynthesis of Protein (Russ.) 241 A . I . SAYKINA

H . - P . K L E B E R , R . CLAUS, O . A S P E R G E R :

Enzymology of the n-Alkane Oxydation a t Acine-

tobacter (Ger.)

251

C. ANTHONY: Methanol Oxidation and Growth Yields in Methyolotropic Bacteria: A Review (Eng.) 261 Y. A.

TROTSENKO

: Metabolic Features of Methane- and Methanol-Utilizing Bacteria (Eng.) 269

H . J . R E H M , L . HORTMANN, I . R E I F F :

Regulation of t h e Formation of F a t t y Acids a t t h e

Microbial Oxidation of Alkanes (Ger.)

279

U. ISKE, H.-J. ROST, S. PORTIUS: A Laboratory Fermentor System for Basic Research in Biotechnology. I. Machine E q u i p m e n t (Ger.) 289 D . C. DAN, K . - I . LINOW, B . PHILIPP, G . SCHULZ, E . W . U N G E R : M o r p h o m e t r i c a l

Analysis

of the Enzymatic Hydrolysis of Cellulose (Ger.)

293

Number 4. 1983 J . C. SENEZ: Developments in t h e Field of Protein Enrichment of Foods and Feeds (Eng.) . 299 T. B. KORONELLI: Penetration of Hydrocarbons into Bacteris Cells (Russ.)

309

O. ASPERGER, R . MÜLLER a n d H . - P . K L E B E R : I s o l a t i o n of C y t o c h r o m P - 4 5 0 a n d t h e c o r r e s -

ponding Reductase System f r o m Acinetobacter (Ger.)

319

and A. T A N A K A : Peroxisomes of Alkane-Utilizing Yeasts Methabolic Functions and Practical Aspects (Eng.) 327 S. FUKUI

J . P . VANDECASTEELE,

D . BLANCHET,

J . P . TASSIN,

A . M . BONAMY

and

L . GUERRILLOT:

Enzymology of Alkane Degradation in Pseudomonas aeruginosa (Eng.)

339

K . S A T T L E R and L. W Ü N S C H E : Possibilities of Obtaining Coupled Products of the Microbial SCP Production on Basis of Hydrocarbons (Ger.) 345 W . BRENDLER,

J . BAUCH,

G. A. LÜBBERT,

L. WÜNSCHE,

R . HEDLICH,

K . TRIEMS,

E. N.

SHDANNIKOWA: Special Aspects of Nonsteril Yeast Production on Basis of Hydrocarbons (Ger.) 351 J . D. S C H N E I D E R , E. F A U L H A B E R and M. R I N G P F E I L : Influence of t h e Ions Cu 2+ , Zn 2 + and Mn2+ on Macromolecular Composition of the Yeast Candida utilis M 72 (Ger.) 357 V. A. VAVILIN: Criterion for Aerobic Biological Treatment Design (Eng.)

363

U. L Ö H S and tion by Acinetobacter calcoaceticus (Ger.)

371

D . HAFERBURG, O. ASPERGER,

H.-P. KLEBER:

Regulation of t h e Alkan Utiliza-

J . ENGEL, A . STEUDEL a n d K . SATTLER: I n v e s t i g a t i o n s o n G e n e t i c of M e t h y l o t r o p h i c B a c -

teria (Ger.)

375

H.-P. S C H M A U D E R and D. G R Ö G E R : Selection and Characteristics of potential Alkaloid Producing Strains of Claviceps (Ger.) 379 1*

"Acta Biotechnologica" publishes reviews, original papers, short communications and reports out of the whole area of biotechnology. The journal shall promote the foundation of biotechnology as a new, homogeneous scientific field. According to biotechnology are microbial technology, biochemical technology and technology of synthesyzing and applying of bioanalogous reaction systems. The technological character of the journal is guarenteed thereby that microbial, biochemical, chemical and physical contributions must show definitely the technological relation.

Terms of subscription for the journal "Acta Biotechnologica" Orders can be sent — in the GDR: to Postzeitungsvertrieb or to Akademie-Verlag, DDR - 1086 Berlin, Leipziger Str. 3 - 4 ; — in the other socialist countries: to a book-shop for foreign languages literature or to the competent news-distributing agency; — in the FRG and Berlin (West): to a book-shop or to the wholesale distributing agency Kunst und Wissen, Erich Bieber OHG, Wilhelmstr. 4—6, D - 7000 Stuttgart 1; — in the other Western European countries: to Kunst und Wissen, Erich Bieber GmbH, Dufourstr. 51, CH - 8008 Zürich; — in other countries: to the international book- and journal-selling trade, to Buchexport, Volkseigener Außenhandelsbetrieb der DDR, DDR - 7010 Leipzig, Postfach 160, or to Akademie-Verlag, DDR - 1086 Berlin, Leipziger Str. 3—4.

Acta Biotechnologica Herausgeber: Institut für technische Chemie der AdW, DDR - 7050 Leipzig, Permoserstr. 15 (Direktor: Prof. Dr. Manfred Ringpfeil) und Institut für Technische Mikrobiologie DDR - 1017 Berlin; Alt-Stralau 62 (Direktor: Dipl. Ing. G. Vetterlein). Verlag: Akademie-Verlag, DDR - 1086 Berlin, Leipziger Straße 3—4; Fernruf: 2236221 und 2236229; Telex-Nr.: 114420; Bank: Staatsbank der DDR, Berlin, Konto-Nr.: 6836-26-20712. Redaktion: Dr. Lothar Dimter (Chefredakteur), Martina Bechstedt (Redakteur), DDR-7050 Leipzig, Permoserstr. 15; Tel.: 2392255. Veröffentlicht unter der Lizenznummer 1671 des Presseamtes beim Vorsitzenden des Ministerrates der Deutschen Demokratischen Republik. Gesamtherstellung: VEB Druckhaus „Maxim Gorki", DDR - 7400 Altenburg. Erscheinungsweise: Die Zeitschrift „Acta Biotechnologica" erscheint jährlich in einem Band mit 4 Heften. Bezugspreis eines Bandes 120,— M zuzüglich Versandspesen; Preis je Heft 30,— M. Bestellnummer: 1094/3. Urheberrecht: Alle Rechte vorbehalten, insbesondere der Übersetzung. Kein Teil dieser Zeitschrift darf in irgendeiner Form — durch Photokopie, Mikrofilm oder irgendein anderes Verfahren — ohne schriftliche Genehmigung des Verlages reproduziert werden. — All rights reserved (including those of translations into foreign languages). No part of this issue may be reproduced in any form, by photoprint, microfilm or any other means, without written permission from the publishers. © 1983 by Akademie-Verlag Berlin. Printed in the German Democratic Republic. AN (EDV) 42133

Acä BlotedNtoiica Journal of microbial, biochemical and bioanalogous technology

Volume 3

Edited a t Institute of Technical Chemistry of the Academy of Sciences of the G.D.R.; Leipzig and Institute of Technical Microbiology; Berlin by M. Ringpfeil, Leipzig and G. Vetterlein, Berlin

Editorial board: M. E. Beker, Riga H. W. Blanch, Berkeley S. Fukui, Kyoto H. G. Gyllenberg, Helsinki G. Hamer, Zurich J . Hollo, Budapest M. V. Iwanow, Pushchino F. J u n g , Berlin H. W. D. Katinger, Vienna K. A. K a l u n j a n z , Moscow J . M. Lebeault, Compiegne D. Meyer, Leipzig

P. Moschinski, Lodz A. Moser, Graz M. D. Nicu, Bukarest Chr. Panajotov, Sofia L. D. Phai, Hanoi H. Sahm, Jülich W. Scheler, Berlin R . Schulze, K o t h e n B. Sikyta, Prague G. K . Skrjabin, Moscow M. A. Urrutia, H a b a n a J . E. Zajic, El Paso

Redaction:

L. Dimter, Leipzig

1983 Number 4

A K A D E M I E - V E R L A G

•

B E R L I N

"Acta Biotechnologica" publishes reviews, original papers, short communications and reports out of the whole area of biotechnology. The journal shall promote the foundation of biotechnology as a new, homogeneous scientific field. According to biotechnology are microbial technology, biochemical technology and technology of synthesyzing and applying of bioanalogous reaction systems. The technological character of the journal is guaranteed thereby t h a t microbial, biochemical, chemical and physical contributions must show definitely the technological relation.

Terms of subscription for the journal "Acta Biotechnologica" Orders can be sent — in the GDR: to Postzeitungsvertrieb or to Akademie-Verlag, DDR-1086 Berlin, Leipziger Straße 3 - 4 ; — in the other socialist countries: to a book-shop for foreign languages literature or to the competent news-distributing agency; — in the FKG and Berlin (West): to a book-shop or to the wholesale distributing agency Kunst und Wissen, Erich Bieber, Wilhelmstr. 4—6, D-7000 Stuttgart 1; — in the other Western European countries: to Kunst und Wissen, Erich Bieber GmbH. Dufourstraße 51, Ch-8008 Zürich; — in other countries: to the international book- and journal-selling trade, to Buchexport, Volkseigener Außenhandelsbetrieb der D D R , DDR-7010 Leipzig, Postfach 160, or to Akademie-Verlag, DDR-1086 Berlin, Leipziger Straße 3—4. Acta Biotechnologica Herausgeber: Institut für technische Chemie der AdW, DDR-7050 Leipzig, Permoserstr. 15 (Direktor: Prof. Dr. Manfred Ringpfeil) und Institut für Technische Mikrobiologie DDR-1017 Berlin; Alt-Stralau 62 (Direktor: Dipl. Ing. G. Vetterlein) Verlag: Akademie-Verlag, DDR-1086 Berlin, Leipziger Straße 3—4; "Fernruf: 2 23 6221 und 2 23 62 29; Telex-Nr.: 114420; Bank: Staatsbank der DDR, Berlin, KontoNr.: 6836-26-20712. Redaktion: Dr. Lothar Dimter (Chefredakteur), Martina Bechstedt (Redakteur), DDR-7050 Leipzig, Permoserstr. 15; Tel.: 2392255. Veröffentlicht unter der Lizenznummer 1671 des Presseamtes beim Vorsitzenden des Ministerrates der Deutschen Demokratischen Republik. Gesamtherstellung: V E B Druckhaus „Maxim Gorki" DDR-7400 Altenburg. Erscheinungsweise: Die Zeitschrift „Acta Biotechnologica" erscheint jährlich in einem Band mit 4 Heften. Bezugspreis eines Bandes 120,— M zuzüglich Versandspesen; Preis je Heft 30,— M. Bestellnummer dieses Heftes: 1094/3/4. Urheberrecht: Alle Rechte vorbehalten, insbesondere der Übersetzung. Kein Teil dieser Zeitschrift darf in irgendeiner Form — durch Photokopie, Mikrofilm oder irgendein anderes Verfahren — ohne schriftliche Genehmigung des Verlages reproduziert werden. — All rights reserved (including those of translations into foreign languages). No part of this issue may be reproduced in any form, by photprint, microfilm or any other means, without written permission from the publishers. © 1983 by Akademie-Verlag Berlin. Printed in the German Democratic Republic. AN (EDV) 42133

Acta Biotechnol. 3 (1983) 4, 299— 308

New Developments in the Field of Protein Enrichment of Foods and Feeds (P.E.F.F) J . C. SENEZ

CNRS/LCB, 31, Chemin Joseph-Aiguier, 13009 Marseille, Prance Paper given at the Leipzig Symposium on Biotechnology 1982, Leipzig 2 1 . - 2 5 . 9. 1982

Summary U p to the year 2000 the total demand of protein for animal feeding will rise by 146%. This demand can not be covered by agriculture alone. According to FAO data it will be an expected deficit of some 400 million tons of cereals at this time all over the world. Therefore the large scale industrial production of SCP from alkanes, methanol and other raw materials will undoubtely become soon an absolute necessity. On the other hand, a new process of solid state fermentation (ORSTOM/IRCHA process) having special interest for the developing countries has been recently set-up in France. Based on dried and crushed potatoes or cassava (manioc or tapioca) protein enrichment by fermentation of feeds or foods (PEFF) in a simple manner without aseptic conditions with the filamentous fungus Aspergillus hennebergii is carried out. The process is presently optimized and scaled-up to the size of a 700 kg (dry weight) fermenter. Nutrititional and toxicological assays have demonstrated complete safety and excellent nutrititional value of the product, which is applicable as a feedstuff for breading of e.g. pigs or chickens.

Zusammenfassung Der Proteinbedarf für die Tierfütterung wird sich im Weltmaßstab bis zum Jahre 2000 um 146% erhöhen und kann nicht allein durch die Landwirtschaft (Pflanzenproduktion) gedeckt werden. Laut Angabe der FAO ist dann ein Defizit von 400 Millionen Tonnen Getreide zu erwarten. Deshalb wird die industrielle Großproduktion von SCP aus Alkanen, Methanol und anderen Rohstoffen zweifellos zur Notwendigkeit. I n Frankreich wurde ein Feststoff-Fermentationsprozeß (ORSTOM/IRCHA-Prozeß) entwickelt, der besonders in Entwicklungsländern auf village-level-Basis betrieben werden kann. Getrocknete und gemahlene Kartoffeln oder Cassava (Maniok oder Tapioca) werden durch Fermentation mit dem Mycelpilz Aspergillus hennebergii in einem Einfach-Verfahren ohne sterile Bedingungen mit Protein angereichert. Der Prozeß wurde bis zum 700 kg (Trockengewicht)-Maßstab optimiert und wird gegenwärtig in verschiedenen Ländern wie Brasilien, Indien, Indonesien u. a. in noch größerem Maßstab erprobt. Fütterungs- und toxikologische Untersuchungen erbrachten den Nachweis der vollständigen Unbedenklichkeit und des hohen ernährungsphysiologischen Wertes des Produktes als Futtermittel für die Schweinemast, die Hühneraufzucht u. a.

Before reporting on protein enrichement of feeds and foods by fermentation [PEFF) it is necessary to formulate some general remarks on the protein problem in the world and on the possible contributions of biotechnology to its solution are formulated. 1*

300

J . C. SENEZ

I n spite of economical and political difficulties, large scale industrial production of SCP's from alkanes, methanol and other raw materials will undoubtedly become soon a n absolute necessity. The protein demand for direct h u m a n consumption and animal feeding will inevitably increase in the course of the two coming decades, due to both demographic growth and economical development. As shown b y Table 1, the world population will reach 6.5 billion in the year 2000 and most of this increase will take place in t h e already under- fed parts of the Third World. Therfore, only in order to keep the alimentary standards at their present widely unsufficient level, the agricultural production of food should increase b y 45 per cent. Table 1. World demographic and economical trends 1 Population 2

GDP 3

1970

1980

2000

A%/year

1970

1980

2000

A%/year

Industrialized 1108 countries

1220

1441

0.9

2458

3458

6446

3.2

Developing countries Group I 4 Group II 5

357 2154

476 2704

856 4104

2.9 2.2

280 185

500 244

1632 628

5.9 4.1

World

3620

4400

6405

2.0

890

1165

2071

2.8

'} 2 ) 3 ) 4 )

data from U . N . : The Future of World Economy, Oxford Univ. Press. 1977 millions Gross Domestic Product, in U S dollars (1970) per capita countries with large potential ressources in Middle East and Africa (oil producing), Latin America, Africa and Asia (tropical). 6 ) countries with low potential ressources in Africa (arid), Latin America and Asia

During the same period, the overall G D P is expected to rise twice faster t h a n population. As demonstrated b y K. HOSHIAI [3], this will inevitably result in greater consumption of animal protein and larger demand of protein for animal feeding (Fig. 1.). F r o m these considerations, one should estimate t h a t from now to the year 2000, the total demand of cereals for human consumption and animal feeding will rise by 75% and the demand of protein for animal feeding by 146% (Table 2). Such demand can not be expected to be covered b y agriculture alone. According to FAO data, the average world production of cereals in the past two decades has increased only b y 2.8% per year, corresponding to an expected deficit [5] of some 400 million tons in the year 2000. Similarly, the deficit of protein for animal feeding will be in between 65 and 120 million tons depending if the agricultural production is to progress exponentially or, as it is more probable, linearly. An other important and maybe decisive argument for development of the SCP industry is the obvious necessity for the industrialized countries importing proteins for animal feeding to escape from the corresponding currency drain and from the menace of economical and political pressure exerted by the exporting countries. A priori, one could also expect the SCP industry to provide a decisive contribution to the problem of hunger in the Third World. I n this regard, however, there are several major obstacles. To be economically viable, a SCP production unit should have a minimal capacity of a t least 100,000 tons per year, corresponding to a capital cost of 50 to 70 million dollars. On the other hand, a plant producing 100,000 tons of SCP from paraffins would require

Acta BiitMiuUiica Journal of microbial, biochemical and bioanalogous technology

Volume 3 • 1983

AKADEMIE-VERLAG

Edited at Institute of Technical Chemistry of the Academy of Sciences of the G.D.R.; Leipzig and Institute of Technical Microbiology; Berlin by M. Ringpfeil, Leipzig and G. Vetterlein, Berlin

Editorial board: M. E. Beker, Riga H. W. Blanch, Berkeley S. Fukui, Kyoto H. G. Gyllenberg, Helsinki G. Hamer, Zurich J . Hollo, Budapest M. V. Iwanow, Pushchino F. Jung, Berlin H. W. D. Katinger, Vienna K. A. Kalunjanz, Moscow J . M. Lebeault, Compiegne D. Meyer, Leipzig

P. Moschinski, Lodz A. Moser, Graz M. D. Nicu, Bukarest Chr. Panajotov, Sofia L. D. Phai, Hanoi H. Sahm, Jülich W. Scheler, Berlin R. Schulze, Kothen B. Sikyta, Prague G. K. Skrjabin, Moscow M. A. Urrutia, Habana J . E. Zajic, El Paso

Redaction:

L. Dimter, Leipzig

•

BERLIN

2

Contents of Volume 3

Contents of Volume 3 (1983) Number 1 . 1 9 8 3 K. H.

STEINKRAUS

: Traditional Food Fermentations as Industrial Resources (Eng.) . . . .

3

and M. J E C H O R E K : Growth Characteristics of a Thermotolerant Strain of Lodderornyces elongisporus grown on Sucrose (Eng.)

J . D . SCHNEID B R , R . H A N S E L ,

R . HEDLICH

K. H. H OFM ANN and W. BABEL: Methylglyoxal—a toxic Fermentation Product (Ger.) . . .

13 21

P. L A I T E : Changes of the Genetic Patterns and Microevolutional Processes under the Conditions of an intensive Lysine Production at some Strains of Brevibacterium (Russ.) . . . .

27

and P. W E L O N O W : Effect of the Chemical Insecticide Fekamatribufon EC 50 on Acricultural Pesticides obtained from Bacillus thuringiensis (Ger.) . .

33

and H . R U T T L O F F : Production and Characterization of Provulgaris (Ger.)

39

Z. L I B U D Z I S Z , H. OBERMAN and K. M A L I K : Overproduction of Aminoacids and the Activity of Methanol Dehydrogenase of Protaminobacter thiaminophagus Mutants (Eng.)

49

A. OPOKTF and Amylase (Eng.)

53

M.

S. ILIEWA,

I W . STOJANOW

A . LEUCHTENBERGER, A . TÄUFEL

teases of Thermoactinomyces

N . URAIH:

Saccharification of Cassava (Manihot esculenta) with Millet

E. W. J W A N N Y and M. M. R A S H A D : Sugar Compositions of Extra- and Intracellular Polysaccharides produced by Candida lipolytica (4124) grown on n-Hexadecane (Eng.)

59

V. W. OGUNDERO : Factors affecting Growth and Cellulose Hydrolysis by the Thermotolerant Aspergillus nidulans from Composts.(Eng.)

65

S. KONÉ and U. BEHRENS: The Maintenance Metabolism and the "maximum" Yield Value (Ger.)

73

S.

76

KONÉ

and

U. BEHRENS:

Meaning of the True Maximum Specific Growth Rate (Ger.) .

.

A. SCHMIDT, D . KUCHENBECKER a n d H . DÄBERITZ: P r e l i m i n a r y R e s u l t s in t h e F l u o r o m e t r i c

Investigation of Fermentation Processes (Eng.)

79

P. S T O L L and F. GLOMBITZA : Heat Flow Measurements in Aerobic Microbial Growth Processes with a Nonisothermal Calorimeter operating directly (Eng.) . . . .

83

and R . D . R E C K N A G E L : A new Biometrical Screening Method as applied to Nutrient Optimization in Fermentation (Eng.)

89

B . HEINRITZ, H . BOOKER

H . - P . SCHMAUDER, D . GRÖBER and W . BADE: Construction of B i o r e a c t o r s suited for the

Cultivation of Plant Cells (Ger.)

93

Number 2 . 1 9 8 3 E . GARA Y , E . A L C A I D E and Physiology and Industrial Applications (Eng.)

J.P.MARTÍNEZ,

E.HERNÁNDEZ:

The Genus

Thiobacillus:

99

E. S T I C H E L and T H . B L E Y : Use of Biorhythmic Processes for Increasing the Efficiency of Carbon-Compound Conversion by Microorganisms (Eng.) 125 B . HEINRITZ, G . ROGGE,

H . LTJTHER, A . CLAUS, W . B Ü T T N E R

Yeast Cells (Ger.)

and

E . SCHUSTER:

Separation of mechanically disrupted

133

M. BULLMANN, G . - J . U H L E N H U T and G . S C H I N D L E R : Kinetics of Mixed Substrates at Citric Acid Accumulation by Saccharomycopsis lipolytica EH 59 (Ger.) . 143 U . ISKE,

CH. G W E N N E R ,

and I. M. C H I R K O V : Electrophysical Methods of Monitoring Biosynthesis (Eng.) 1 5 5 E. DANIELOVÁ, V . F A R K A S and S. B A U E R : Enzymatic Hydrolysis and Ethanol Fermentation of Lignocellulosic Materials (Ger.) 163 N. V . SEDYKH

A. SCHMIDT : The Technological Possibilities for Measuring of Physiological Population State Features in Fermentation Practice (Eng.) 171

Contents of Volume 3

3

dinh P H A I : Biochemical and Microbiological Aspects of the "Nuoc-mam" Industry in Vietnam (Ger.) 177 LE

and H . P . K L E B E R : Regulation of the extracellular Lipase from an Alkan Utilizing Strain of Acinetobacter (Ger.) 185 D . HAFERBURG

Number 3 . 1 9 8 3 A.

MOSER:

Formal Macroapproach to Bioprocessing-Modeling with Analogies (Eng.) . . . .

195

S . K. S E N , M. C H A T T E R J E E , S . P. C H A T T E R J E E , A . K. B A N E R J E E : Activity of Diaminopimelic Acid Decarboxylase in Bacteria Producing Lysine (Eng.) 217 E . E M A N U I L O Y A , A. K A I M A K T C H I E V : Production of Neutral and Alkaline Proteases in Batch and Chemostat Cultures by Bacillus mesentericus 76 (Eng.) 221 M. R I N G P F E I L : SCP-Production on the Basis of Hydrocarbons (Ger.) 227 N . B . GRADOVA, E . M . DIKANSKAYA, S . N . ROBYCHEVA, G . S . RODIONOVA, M . B . BITTEYEVA,

A. I. S A Y K I N A : Characteristics of Yeast Strains for the Production of SCP on the Basis of Petroleum Hydrocarbons. Specific Growth Properties and Biosynthesis of Protein (Russ.) H.-P. K L E B E R , R . CLAUS, 0 . A S P E R G E R : Enzymology of the n-AIkane Oxydation at Acinetobacter (Ger.) C. A N T H O N Y : Methanol Oxidation and Growth Yields in Methyolotropic Bacteria: AReview (Eng.) Y. A. T R O T S E N K O : Metabolic Features of Methane- and Methanol-Utilizing Bacteria (Eng.) H . J . R E H M , L . H O R T M A N N , I . R E I F F : Regulation of the Formation of Fatty Acids at the Microbial Oxidation of Alkanes (Ger.) U. I S K E , H.-J. R O S T , S. P O R T I U S : A Laboratory Fermentor System for Basic Research in Biotechnology. I. Machine Equipment (Ger.) D . C . D A N , K . - I . L I N O W , B . P H I L I P P , G . S C H U L Z , E . W . U N G E R : Morphometrical Analysis of the Enzymatic Hydrolysis of Cellulose (Ger.)

241 251 261 269 279 289 293

Number 4. 1983 J . C. S E N E Z : Developments in the Field of Protein Enrichment of Foods and Feeds (Eng.) . 299 T . B. K O R O N E L L I : Penetration of Hydrocarbons into Bacteris Cells (Russ.) 309 O . A S P E R G E R , R. M Ü L L E R and H.-P. K L E B E R : Isolation of Cytochrom P - 4 5 0 and the corresponding Reductase System from Acinetobacter (Ger.) 319 S. F U K U I and A. T A N A K A : Peroxisomes of Alkane-Utilizing Yeasts Methabolic Functions and Practical Aspects (Eng.) 327 J . P . VANDECASTEELE,

D . BLANCHET,

J . P . TASSIN,

A . M . BONAMY

and

L . GUERRILLOT:

Enzymology of Alkane Degradation in Pseudomonas aeruginosa (Eng.) 339 K . S A T T L E R and L . W Ü N S C H E : Possibilities of Obtaining Coupled Products of the Microbial SCP Production on Basis of Hydrocarbons (Ger.) 345 W . BRENDLER,

J . BAUCH,

G. A. LÜBBERT,

L . WÜNSCHE,

R . HEDLICH,

K . TRIEMS,

E. N.

Special Aspects of Nonsteril Yeast Production on Basis of Hydrocarbons (Ger.) , J . D. S C H N E I D E R , E. F A U L H A B E R and M. R I N G P F E I L : Influence of the Ions Cu2+, Zn2+ and Mn2+ on Macromolecular Composition of the Yeast Candida utilis M 72 (Ger.) V . A. V A V I L I N : Criterion for Aerobic Biological Treatment Design (Eng.) D . H A F E R B U R G , O . A S P E R G E R , U. L Ö H S and H . - P . K L E B E R : Regulation of the Alkan Utilization by Acinetobacter calcoaceticus (Ger.) J . E N G E L , A. S T E U D E L and K. S A T T L E R : Investigations on Genetic of Methylotrophic Bacteria (Ger.) • H.-P. S C H M A U D E R and D. G R Ö G E R : Selection and Characteristics of potential Alkaloid Producing Strains of Claviceps (Ger.) SHDANNIKOWA:

1*

351 357 363 371 375 379

"Acta Biotechnologica" publishes reviews, original papers, short communications and reports out of the whole area of biotechnology. The journal shall promote the foundation of biotechnology as a new, homogeneous scientific field. According to biotechnology are microbial technology, biochemical technology and technology of synthesyzing and applying of bioanalogous reaction systems. The technological character of the journal is guarenteed thereby that microbial, biochemical, chemical and physical contributions must show definitely the technological relation.

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Acta Biotechnologica Herausgeber: Institut für technische Chemie der AdW, DDR - 7050 Leipzig, Permoserstr. 15 (Direktor: Prof. Dr. Manfred Ringpfeil) und Institut für Technische Mikrobiologie DDR - 1017 Berlin; Alt-Stralau 62 (Direktor: Dipl. Ing. G. Vetterlein). Verlag: Akademie-Verlag, DDR - 1086 Berlin, Leipziger Straße 3—4; Fernruf: 2236221 und 2236229; Telex-Nr.: 114420; Bank: Staatsbank der DDR, Berlin, Konto-Nr.: 6836-26-20712. Redaktion: Dr. Lothar Dimter (Chefredakteur), Martina Bechstedt (Redakteur), DDR-7050 Leipzig, Permoserstr. 15; Tel.: 23 92255. Veröffentlicht unter der Lizenznummer 1671 des Presseamtes beim Vorsitzenden des Ministerrates der Deutschen Demokratischen Republik. Gesamtherstellung: VEB Druckhaus „Maxim Gorki", DDR - 7400 Altenburg. Erscheinungsweise: Die Zeitschrift „Acta Biotechnologica" erscheint jährlich in einem Band mit 4 Heften. Bezugspreis eines Bandes 120,— M zuzüglich Versandspesen; Preis je Heft 30,— M. Bestellnummer: 1094/3. Urheberrecht: Alle Rechte vorbehalten, insbesondere der Übersetzung. Kein Teil dieser Zeitschrift darf in irgendeiner Form — durch Photokopie, Mikrofilm oder irgendein anderes Verfahren — ohne schriftliche Genehmigung des Verlages reproduziert werden. — All rights reserved (including those of translations into foreign languages). No part of this issue may be reproduced in any form, by photoprint, microfilm or any other means, without written permission from the publishers. © 1983 by Akademie-Verlag Berlin. Printed in the German Democratic Republic. AN (EDV) 42133

301

Protein Enrichment Animal Protein

10.000

Tota! Protein

8,000 6,000 5,000 4,000

¿0

30 40 50 60 70 80 30 Protein Supplies per Doy per Capita fgj

correlation Unes in 1972 -74 average for reference Pig. 1. Correlation between gross domestic product (GDP) and per capita consumption of total and animal protein. (From K . HOSHIAI, ref. [2])

an equal supply of substrate and should thus be associated with an oil refinery having a minimal capacity of about 3 to 5 million tons of crude per year. Similar considerations apply to the production of SCP from natural gas or from methanol. Such facilities are obviously absent in most non oil-producing countries of Asia, Africa and Latin America. Moreover, these countries may not have a potential market nor an appropriate transportation and distribution network for the commercialization of 100,000 tons SCP per year. Clearly, those countries which can not presently import food or feeds because of currency shortage will also not be able to import industrial SCP-s from abroad. Consequently, it is of utmost importance for them to develop their own protein resources. Besides hydrocarbons and methanol a wide variety of raw materials potentially utilizable for SCP production might be considered. However most of them are available at too high a cost to be economically competitive or in quantities too low for protein production on a really significant scale. Amongst the substrates at suitable cost and supply, special emphasis is usually put on lignocellulosic materials, but, at the moment, the many attempts made in this direction have had little success, the main difficulty being the lack of cellulolytic organisms with a adequate growth rate and yield.

302

J . C. SENEZ

Table 2. Prospective evolution of world supplies for animal feeding A/year

1970

1980

1990

2000

3.3% 2.8%

1027 1027

1393 1360 33

1946 1800 146

2804 2381 423

Cereals1 total demand 2 production deficit otein rich feeds

—

3

demand production

4.5%

62.5

98.0

153.9

241.5

4.5%

36.4 26.1 62.5

57.0 29.1 86.1 11.9

89.5 32.5 122.0 31.9

140.4 36.3 176.7 64.8

36.4 26.1 62.5

52.2 29.1 81.3 16.7

68.0 32.1 100.1 53.8

83.8 35.1 118.9 122.6

exponential progression — — — —

soymeal other meals total deficit

1-1% 3.5% -

linear progression — — — —

soymeal other meals total deficit

1.58 Mt 0.3 Mt 1.88 Mt —

) million metric tons (Mt) ) human consumption included 3 ) million metric tons (Mt) of soymeal equivalent ( 4 4 % protein) 1

2

In contrast, starchy materials, and more specifically cassava in the tropical regions or potatoes in more temperate climates, are of obvious interest, due to both high productivity per hectare, and an excellent rate of conversion to biomass by a great number of fast growing microorganisms. There is in the literature a considerable number of publications in SCP production from starchy substrates. However, most of these studies have only been made at the laboratory scale, without enough consideration for economical feasibility. This is namely the case of processes involving classical fermentation in liquid medium, under aseptic conditions, followed by biomass separation and drying. As for SCP production from paraffins or methanol, the optimization of such sophisticated technology would require a minimal production well over the possibilities of most developing countries and would involve high investment and operation costs. Moreover in the developing countries the collection, transportation and storage of large quantities of raw materials would result in major difficulties. In order to overcome these difficulties, several authors have prefered a simpler technology consisting in mere protein enrichment of starchy raw materials by aerobic fermentation. Some of these processes using amylolytic yeasts or filamentous fungi have been developed at the pilot-plant scale. However, most attempts, if not all of them, were designed for large production units and for cultivation of the microorganisms in liquid medium, thus requesting a costly final concentration of the biomass. To be economically competitive, the bioconversion of starch into protein should be performed by a procedure workable at the rural level, in a system combining side by side the production of the raw material, the fermentation process, and the direct utilization for animal feeding or human consumption. The obvious advantage of such an integrated approach is to suppress transportation problems and to prevent intermediary profit and speculation which would inevitably take place if either the raw material or the product were commercialized.

303

Protein Enrichment

From a technological point of view, a bioconversion process intended for rural application should not require aseptic conditions and should be performed with simple and cheap equipment, in a single operation. On the other hand, the final product must be sufficiently rich in protein to be utilized for animal feeding as such, without a secondary concentration step. This last requisite implicates a difficulty which was responsible for the failure of many previous attempts. I n a mash of starchy material dense enough to be directly utilizable, the major problem is to maintain aerobic conditions and oxygen transfer efficiency so as to prevent anaerobic contamination of the culture. These considerations have led us to set-up in France a new process of solid state fermentation, which was developed since 1976 in collaboration with Drs. M. Raimbault (Office de Recherche Scientifique et Technique Outre-Mer, ORSTOM) and F. Deschamps (Institut National de Recherche en Chimie Appliquée, IRCHA) [3, 4, 6]. SUBSTRATE water 1!.

I

STEAMING 1,3 kg 10 m n - 75 °C water 0,61, mineral-salts spores control humidity pH temperature

DRYING 1,5!. water

PRODUCT 780g; 20% protein 25-30% residual sugars

\ NO EFFLUENT

>

Fig. 2. Principle of the ORSTOM/IRCHA process for protein enrichment by solid-state fermentation.

The ORSTOM/IRCHA process is schematically illustrated by Fig. 2. Dried and crushed potatoes or cassava (syn. manioc or tapioca) containing initially less than 1% protein are introduced in a commercial bakery-kneader adequatly modified for aeration and fermentation control. The first step consists in glutinization of starch by steaming a t 75 °C for 10 min. After cooling, the material is added with water containing a nitrogen source (urea and ammonium sulfate), phosphates, mineral salts and 2 x 107 spores per g of substrate of filamentous fungus (Aspergillus hennebergii). Initial p H is adjusted at 4.5. By gentle mechanical stirring, the inoculated substrate with a water content of 65% takes spontaneously the form of well separated granules of 1 to 2 m m diameter, the mass of which is freely permeable to aeration from the bottom of the tank. The substrate is incubated at 37 °C for 24—26 hours under aeration, with continuous control of temperature, p H and water content. Fig. 3 shows an experimental fermenter of 1.200 litres with a capacity of 200 kg dry weight of fermented material. The monitoring devices are schematically represented on

304

J . C. SENEZ

Fig. 3. Solid-state fermenter (1,200 litres)

Fig. 4. Temperature (32°C) is controlled by aeration from the bottom of the tank, by mechanical stirring, and by water spraying, this last device being also utilized for pH and moisture adjustment. Photographs taken with a scanning electron microscope shows the initial starch granules (Fig. 5 A), the effect of glutinization (Fig. 5 B ) , and the final shape of the product completely colonized by dense fungal mycelium (Fig. 5C).

Pig. 4. Monitoring devices. Control of temperature (thermistance), pH (glasselectrode), and moisture (strain-gauges). Temperature automatically adjusted by aeration, mechanical stirring and water spraying, this last device being also monitored for pH and moisture adjustment.

Protein Enrichment

305

Fig. 5. Electron scanning micrographs. (A) initial aspect of starch granules (cassava flour); (B) glutinized material and germination of fungal spores; (C) 'final aspect after 26 h fermentation.

306

J . C. SENEZ

The kinetics of a culture on cassava are represented by the curves of Fig. 6 showing the consumption of carbohydrate, the production of protein, and the evolution of pH and moisture content. At the end of the culture, i.e. after 26 hours incubation, the final product contains about 20 per cent true protein, measured by the Lowry method, and 25 per cent residual sugar. The fact that the monitorized system of cooling by aeration, stirring and water spraying has to operate for only 5 h over a 26 h incubation time (Fig. 6, curve Q) indicates that the process does consume much less energy than a conventional aerobic fermentation in liquid medium. This point is obviously of great importance for a process to be utilized at the farm level.

Fig. 6. Kinetics of solid-state fermentation. Sugar consumption ( — 0 — ); protein ( 1 ) ; pH ( ); moisture ( ); time of operation of the cooling system ( — 0 — , curve Q).

As demonstrated by M. RAIMBAULT [3], the relatively large quantity of residual sugar is due to the fact that the growth of the organism is limited by the availability of extracellular water, which, at the end of the culture, represents only 20% of the total water content (63%).

The overall protein/carbohydrate conversion factor is 55%. Under the chosen conditions, the sporulation of the mold is completely inhibited. On the other hand, in spite of that the process is non-aseptic, no contamination by anaerobic bacteria takes place, and the initial number of aerobic bacteria decreases by a factor of 10 during the two last doubling times of biomass. This is due to excellent aerobic conditions, to acidic pH, and to the fact that the availability of free water does limit the growth of bacteria well before that of the mold. The results obtained from a variety of raw materials are reported on Table 3. The aminoacid profile of the product is quite satisfactory, with a high lysin content and, in contrast with soybeans and SCPs from yeasts and bacteria, no deficit in methionine and cysteine.

307

Protein Enrichment Table 3. Protein enrichment by solid-state fermentation substrate 1 protein 2

product carbohydrate 3

protein

/o cassava banana banana refuses potato potato wastes from fecula plant

carbohydrate /o

2.5 6.4 6.5 5 5

18-20 20 17

90

80 72 90 65

20 18

25-30 25 33 35 28

!) dry flour : 100 g, water 1 0 0 - 1 2 0 g ; S 0 4 ( N H ) 2 : 9 g; urea : 2.7 g; P 0 4 H K 2 : 5 g. 2 ) determined by LOWRY method 3 ) determined by enzymatic hydrolysis (amyloglucosidase) and SOMOGYI method.

Nutritional and toxicological assays on rats and chicken have demonstrated complete safety and excellent nutritional value, with a protein efficiency and digestibility similar to that of casein. The organism routinely utilized is a strain of Aspergillus hennebergii isolated in Senegal from traditionally fermented cassava food. This strain was selected for optimal amylolytic activity, growth rate and biomass yield. However, similar results have been also obtained with a number of other amylolytic molds (Asp. orizae, and various Rhizopus and Penicillum species) isolated from tempeh, koji and other well known fermented foods of South-East Asia. Table 4. Comparative productivity of protein-rich feeds and protein enrichment by solid-state fermentation Yield ton/ha

Protein content

ton/ha

/o soybean rapeseed sunflower horsebean protein enriched cassava or potato 1

1.8 3.0 2.5 3.2 15.2

34 23.3 22 28 20

0.6 0.7 .0.55 0.9 1.7

yield : 40 tons/ha; 6 2 % humidity; conversion factor (protein/carbohydrate) : 5 5 %

The agro-economical prospects are illustrated by Table 4, showing that one can obtain via solid-state fermentation of cassava or potato about 3 times as much protein per hectare than by cultivating soybean, or other protein-rich pulses. Another interesting estimation, is that one hectare of cassava or potato may provide enough protein for the breeding of some 75 pigs per year (Table 5). The ORSTOM/IRCHA process is actively developed in collaboration with a French firm of bioengineering (SPEICHIM). The fermentation equipment is presently optimized and scaled-up to the size of a 700 kg (dry weight) fermenter, i.e. a capacity sufficient for the breading of approximatively 1400 pigs or 15000 chickens per year.

308

J. C. SENEZ Table 5. Agro-economical prospects of protein enrichment by solid-state fermentation A . P r o d u c t i v i t y of raw material and protein cassava (tons/ha) moisture ( % ) conversion factor (protein/carbohydrate) protein (tons per hectare)

40 62 0.55 1.7

B. Conversion to animal product (pig) 1 alimentary conversion rate protein consumption — birth to weaning 2 — weaning to slaughter 3 — total 4

3:1 11.3 kg 25.5 kg 36.8 kg

C. Overall prospect one hectare of cassava can provide via solid state fermentation the quantitity of protein required f o r breeding 50 pigs x)

data f r o m C. A . SHACKLADY, in " P r o t e i n s f r o m hydrocarbons", Academic N e w Y o r k , 1972 : 1 1 5 - 1 2 8 ; 2 ) birth t o weaning : 70 d a y s ; + 25 k g ; diet with 1 5 % protein; 3 ) weaning t o slaughter : 130 d a y s ; + 65 k g ; diet with 1 0 % protein; 4 ) total : 200 d a y s ; + 1 1 0 kg.

Press,

On the other hand, negotiations are currently in progress for setting-up international cooperation and full-scale rural experimentation in several countries, among which Brazil, India, Indonesia, Mexico and West Indies. Some of these programmes will include the coupling of protein enrichment with biogas production from animal wastes, in order to make the whole system energetically independent. Protein enrichment by fermentation of feeds or foods ( P E F F ) in rural communities is of obvious economical interest not only for the tropical regions but also for european countries with large production of potatoes. Such supply of protein is not a conflicting and mutually exclusive alternative to large scale industrial production of SCP, but these two biotechnological approaches have quite different and complementary economical prospects and both of them will have a most promising future.

References [1] DESCHAMPS, F., MEYER, F.,PRÉBOIS, J. P . : S y m p . Soc. Fcse Microbiol., Toulouse, (1980), p. 135. [2] HOSHIAI, K . : Chem. E c o n o m y and Engin. R e v . 10 (1978) 1 (Japan). [3] RAIMBAULT, M. : Fermentation en milieu solide, T r a v a u x et Docum. O R S T O M , Paris (1981), p. 291. [4] RAIMBAULT, M., ALAZARD, D . : Europ. J. A p p l . Microbiol. Biotechn. 9 (1980) 199. [5] SENEZ, J. C.: Progrès Scientiv., n° 203 (1979) 5. [6] SENEZ, J. C., RAIMBAULT, M., DESCHAMPS, F . : W o r l d A n i m a l R e v . , n ° 3 5 (1980) 36.

Acta Biotechnol. 8 (1983) 4, 3 0 9 - 3 1 8

IIorjiom,eHne yraeßo^opo^OB T.

B.

SaKTcpiiflMii

KOPOHEJIJIH

MOCKOBCKHH rocyAapcTBeHHHö yHHBepcwTeT. EHOJiorHiecKHü (JiaKyjifcTeT. M o c K B a , JleHHHCKHe r o p t i ,

CCCP

Paper given a t t h e Leipzig Symposium on Biotechnology 1982, Leipzig 2 1 . - 2 5 . 9. 1982

Summary I t is demonstrated t h a t aliphatic hydrocarbons (alkanes) penetrate into bacteria cells by t h e way of passive diffusion. The mechanism of this process is different for several bacteria species. A hydrophobic cell wall is essential for t h a t process. I n saprophytic Mycobacteria hydrocarbons are solubilized in the thick hydrophobic cell wall. During the process of absorption hydrocarbons pass through the whole cell wall u p to the membrane. I n the case of Arthrobacteria the hydrocarbons might pass not through t h e whole cell wall, b u t through special lipophilic canals. Mobile hydrocarbon-oxidizing bacteria g. Pseudomonas form peptidoglycolipid and excrete it into the medium. The peptidoglycolipid emulsifies hydrocarbon substrate.

Zusammenfassung E s wird demonstriert, daß die Aufnahme von aliphatischen Kohlenwasserstoffen (Paraffinen) durch Bakterienzellen mittels passiver Diffusion erfolgt. Der Mechanismus dieses Prozesses weist bei verschiedenen Bakterienspecies Unterschiede auf. Voraussetzung f ü r diesen Prozeß ist eine hydrophobe Zellwand. Bei den saprophytischen Mycobakterien werden Kohlenwasserstoffe in der dicken lipophylen Zellwand solubilisiert. Die Aufnahme erfolgt mittels Diffusion durch die Zellwand zur Membran. Bei Arthrobakterien erfolgt die Kohlenwasserstoffaufnahme anscheinend nicht durch die gesamte Zellwand, sondern durch besondere lipophile Kanäle. Die beweglichen kohlenwasserstoffoxidierenden Bakterien der Gattung Pseudomonas bilden ein Peptid-Glycolipid, welches das Kohlenwasserstoff-Substrat emulgiert, und scheiden PeptidGlycolipid ins Medium ab. yrjieBonopoflOKHCJiHiomHeSaKTepnn nrpaiOT orpoMHyio aKonornnecKyio p o j i b , p a 3 p y n i a n He3yeMbie Ps. aeruginosa B KaiecTBe ejpiHCTBeHHoro H C T O H HHKa y r j i e p o ^ a H aHeprnn, He BH3HBai0T xeMOTaKCHC, H HanpaBJieHHoe «BHJKeHHe K HHM SaKTepntt HeB03M0JKH0. OflHaKO CMeob nenTHfl0rjiHK0JiHnHj;a, BbmejiHeMoro KjieTKaMM Ps. aeruginosa, c napa