Genetics and Biotechnology of Lactic Acid Bacteria

Transcription

1 Genetics and Biotechnology of Lactic Acid Bacteria

2 Genetics and Biotechnology of Lactic Acid Bacteria Edited by MICHAEL]. GASSON AFRC Institute of Food Research Norwich UK and WILLEM M. DE VOS Department of Biophysical Chemistry Netherlands Institute for Dairy Research (NIZO) and Department of Microbiology Wageningen Agricultural University The Netherlands m SPRINGER-SCIENCE+BUSINESS MEDIA, B.V.

3 First edition Springer Science+Business Media Dordrecht Originally published by Chapman & Hali in 1994 Softcover reprint ofthe hardcover 1 st edition 1994 Typeset in 10/12 pt Times New Roman by ICON Graphic Services, Exeter ISBN ISBN (ebook) DOI / Apart from any fair deal ing for the purposes of research or private study, or criticism or review, as permitted under the UK Copyright Designs and Patents Act, 1988, this publication may not be reproduced, stored, or transmitted, in any form or by any means, without the prior permission in writing of the publishers, or in the case of reprographic reproduction only in accordance with the terms of the licences issued by the Copyright Licensing Agency in the UK, or in accordance with the terms of licences issued by the appropriate Reproduction Rights Organization outside the UK. Enquiries conceming reproduction outside the terms stated here should be sent to the publishers at the Glasgow address printed on this page. The publisher makes no representation, express or implied, with regard to the accuracy of the informat ion contained in this book and cannot accept any legal responsibility or liability for any errors or omissions that may be made. A catalogue record for this book is available from the British Library. Library of Congress Catalog Card Number: Printed on acid-free text paper, manufactured in accordance with ANSI/NISO Z (Permanence of Paper).

4 Preface Over the past decade a dramatic expansion of research interest in lactic acid bacteria has taken place both in academia and industry. This reflects the industrial importance of these bacteria for a wide variety of fermentation processes, and also the interesting genetic and biochemical properties that their more detailed analysis has revealed. We have seen the field grow from a time when there was a mere handful of laboratories interested in the genetics and molecular biology of lactic acid bacteria to the present situation, when such research is prominent in all of the molecular scientific journals. Despite the importance of lactic acid bacteria and the availability of review articles, a book devoted to their genetics and biochemistry is timely. We have attempted to cover both the genetic technology and the genetics of the major industrially important traits. An emphasis on lactococci will be apparent, reflecting the fact that whilst rapid progress in the analysis of all species of lactic acid bacteria is being made, the mesophyllic lactococci are the best characterized. The first two chapters address general aspects of the genetics and molecular biology of lactic acid bacteria. The emphasis of chapter I is on gene transfer processes, including conjugation, transduction and transformation and it also covers the growing range of transposition phenomena that have been recognized in lactic acid bacteria. The exploitation of generalized recombination processes, especially for chromosomal integration and gene replacement techniques, is included and the recent success in constructing genetic and physical maps of the chromosome is covered. Chapter 2 focuses on the more molecular aspects of lactic acid bacterial genetics. It describes the development of gene cloning vector systems based on both plasmids and chromosomal integration. All aspects of gene expression are described in detail, including transcription, translation, regulation and secretion. The most current available vectors for use in lactic acid bacteria are also described. Chapters 3 to 5 deal with three aspects of the lactic acid bacteria that are prominent in their current and potential industrial applications. Chapter 3 describes the proteolytic systems of lactic acid bacteria and includes both the proteinase and peptidase enzymes and their genes, as well as amino-acid and peptide transport systems. Bacteriophages represent one of the most enduring and intractable problems faced by industrial processes involving lactic acid bacteria. A comprehensive description of bacteriophages is included in chapter 4, which also describes the very interesting and varied mechanisms of bacteriophage defence that have evolved in lactic acid bacteria. The latter have already been exploited in the dairy industry and their continuing in-depth analysis should lead to improved strategies to minimize the bacteriophage problem.

5 VI GENETICS AND BIOTECHNOLOGY OF LACTIC ACID BACTERIA Chapter 5 describes the range of antimicrobial proteins that is produced by lactic acid bacteria. Sophisticated analysis of the biosynthesis of both small peptides and lantibiotics such as nisin is described. These various molecules provide the opportunity to use a range of natural antimicrobial agents in food preservation and protection from food poisoning organisms, and they represent a potential application of lactic acid bacteria beyond their established role in food fermentation. Because of the emphasis on lactococci throughout the book, the final chapter is specifically concerned with the important species of Lactobacillus, Leuconostoc and Streptococcus thermophilus, and ensures that good coverage of all lactic acid bacteria is achieved. All authors are established and active researchers in the field of lactic acid bacterial genetics. We hope that, as a result, this book represents an in-depth critical and comprehensive review of what is currently being achieved in the genetics of lactic acid bacteria. The book should appeal to anyone currently active in lactic acid research, but will also serve as an introduction to the molecular genetics of lactic acid bacteria for advanced students of food science and as an introduction to lactic species for geneticists and molecular biologists working outside the field. We wish to acknowledge the support that lactic acid bacteria research in Europe has received from the EC. Most contributors to the book have benefited from support for collaborative research through the BEP, BAP and BRIDGE programmes of the EC, which has greatly facilitated both transnational interaction and scientific progress. MJG WMdeV

6 Contents 1 Gene transfer systems and transposition M. J. GASSON and G. F. FITZGERALD 1.1 Introduction Conjugation Heterologous conjugation systems Homologous conjugation systems Transduction Transduction in Lactococcus 'pp Transduction in Streptococcus thermophilus Transduction in Lactobacillus spp Transformation Transformation systems Protoplast transformation Electrotransformation Transfection Protoplast fusion Transposition Insertion sequences Heterologous transposons Nisin transposons Generalized recombination Integration processes Recombination genes Chromosome mapping Concluding remarks 43 References 44 2 Gene cloning and expression systems in Lactococci 52 W. M. DE VOS and G. F. M. SIMONS 2.1 Introduction Replicative gene cloning S treptoccal plasm id vectors Lactococcal plasmid vectors Plasmid replication and stability Integrative gene cloning Integration vectors Chromosomal gene amplification and stability Integrative gene expression Gene expression signals Vectors for the selection of expression signals Transcription initiation and termination Translation initiation and codon usage Control of gene expression Lactose utilization and its control Heat shock response Negative regulation of transcription Positive regulation of transcription 84

7 viii GENETICS AND BIOTECHNOLOGY OF LACTIC ACID BACTERIA 2.6 Protein secretion Export-signal selection vectors Sec-dependent secretion Sec-independent secretion 2.7 Expression and secretion systems Expression vectors Secretion vectors 2.8 Food grade systems References Bacteriophages and bacteriophage resistance T. R. KLAENHAMMER and G. F. FITZGERALD 3.1 Introduction 3.2 Types and species of bacteriophages Interactions with hosts Temperate phages Classification of phages of lactic acid bacteria Characterization of bacteriophage genomes 3.3 Gene directed bacteriophage resistance in lactic acid bacteria Interference with bacteriophage adsorption Restriction and modification Abortive infection 3.4 Novel phage defense mechanisms 3.5 Genetic strategies to construct phage-insensitive strains 3.6 Conclusions and perspectives References III The proteolytic system of lactic acid bacteria J. KOK and W. M. DE VOS 4.1 Introduction 4.2 Proteinases Biochemical characterization Caseinolytic specificity and classification of proteinases Cloning and expression of proteinase genes Organization of prim and prtp genes Structural characteristics of PrtP Proteinase maturation by PrtM 4.3 Endopeptidases Endopeptidase NisP Intracellular endopeptidases Endopeptidase PepO 4.4 General aminopeptidases Aminopeptidase A (PepA) Pyrrolidonyl carboxylyl peptidase (PCP) Aminopeptidase N (PepN) Aminopeptidase C (PepC) 4.5 Proline-specific peptidases Prolidases Proline iminopeptidase X-prolyl dipeptidyl aminopeptidase (PepXP) 4.6 Oligo- and carboxypeptidases Dipeptidases

8 CONTENTS IX 4.7 Transport of amino acid and peptides Specific amino acid carriers Di- and tripeptide transport system (DptT) Oligopeptide transport system (Opp) 4.8 Cellular localization of proteolytic enzymes 4.9 Engineering of the proteolytic system Construction and use of mutants Engineering of the proteinase PrtP Future perspectives References Bacteriocins of lactic acid bacteria H. M. DODD and M. J. GASSON 5.1 Introduction 5.2 Small heat-stable bacteriocins Genetic analysis Common structural features Bacteriocin translocation Mode of action 5.3 Large heat-labile bacteriocins Helvetican J 5.4 Lantibiotics in lactic acid bacteria Lactocin S Lacticin Nisin structure and biosynthesis Genetic analysis of nisin biosynthetic genes Expression of nisin determinants Nisin immunity resistance Mode of action Protein engineered nisins Applications 5.5 Concluding remarks References Genetic engineering of lactobacilli, leuconostocs and Streptococcus thermophilus A. MERCENIER, P.H. pauwels and B. M. CHASSY 6.1 Introduction 6.2 Overview of taxonomy and health benefits 6.3 Biochemical traits The Lac-PTS The lac operon Xylose metabolism Protein metabolism 6.4 Plasmid biology and cloning vectors Naturally occuring plasmids Insertion sequences Cloning vectors 6.5 Genetic transfer methods Transformation by electroporation Liposome-mediated protoplast transfection, fusion and transformation

9 x GENETICS AND BIOTECHNOLOGY OF LACTIC ACID BACTERIA Conjugation Transduction 6.6 Gene expression in lactobacilli Transcription Translation Regulation of transcription Heterologous gene expression 6.7 Gene expression in S. thermophilus and leuconostocs 6.8 Chromosomal integration of genes 6.9 Phage and phage resistance 6.10 Concluding remarks Dedication and acknowledgements References Index

10 xi Contributors Bruce M. Chassy WilIem M. de Vos Helen M. Dodd Gerald F. Fitzgerald Mike J. Gasson Todd R. Klaenhammer Jan Kok Department of Food Science, University of Illinois, ABLl03, 1302 West Pennsylvania, IL 61801, USA Department of Biophysical Chemistry, Netherlands Institute for Dairy Research, PO Box 20, NL BA, Ede, The Netherlands and also Department of Microbiology, Wageningen Agricultural University, The Netherlands AFRC Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK Department of Food Microbiology, University College, Cork, Ireland AFRC Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK Department of Food Science, Southeast Dairy Foods Research Center, North Carolina State University, North Carolina, USA Department of Genetics, State University of Groninger, Kerklaan 30, 9751, NN Haren, The Netherlands Annick Mercenier Transgene SA, 11 rue Molsheim, Strasbourg, France Peter H. Pouwels Guus F. M. Simons TNO Medical Biological Laboratory, POBox 5815, 2280 HV Rijswijk, The Netherlands KeyGene, POBox 216, 6700 A E Wageningen, The Netherlands