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Stusy on the Aspects of Microbiology and Engineering in Antibiotic Fermentation

Author: JinZhiHua
Tutor: ZuoPeiLin
School: Zhejiang University
Course: Biochemical Engineering
Keywords: antibiotic fermentation rational selection scale-up kinetic model pristinamycin teicoplani ii rifamyci B Streptomyces pristinaespiralis A ctinoplanes wtchornyce/icuc Amzycolatoposis mediterranei
CLC: TQ465
Type: PhD thesis
Year: 2001
Downloads: 893
Quote: 5
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Executive Summary

Strain improvement, scale-up of fermentation processes and kinetics of the three antibiotics (pristinamycin, teicoplanin, and rifamycin 13) were studied. he production medium of pristinamycin by S(reptomyces prisiinaespiralis ATCC 25486 was optimized. The suitable production medium was achieved by single factor experiments and orthogonal experiments. It contains: starch 3%, glucose 2%, soybean meal 2%, peptone 0.5%, fish meal 0.5%, (NH)2S040.15%, MgSO4 0.1%, KI-12P040.04%, CaCO3O.4%, soybean oil 1%, propyl alcohol 0.4%(added at the 18th hour of batch fermentation). The pH value of the medium is 6.0. Carbon catabolite repression and nitrogen catobolite repression was observed in pristinamycin fermentation. Based on the biosynthesis pathway and the metabolic regulation of pristinamycin. teicoplanin and rifamycin B, strain improvement of these antibiotics was performed by mutation and rational screening to improve their productivity and product quality. According to the metabolic pathway of pristinamycin, a rational selection procedure with U. V. mutation was performed to obtained high pristinamycin productivity. A strain S. pristinaespiralis 12-55 with 0.1%AAr, 0.3%AAt, 0.1%Valr, 0.1 %KTMr, and 0.1% DOGr was obtained, whose pristinamycin productivity reaches 3000tt/ml which was 100 times higher than that of the parent strain ATCC 25486. The analysis of kinetic experiments indicated that the pristinamycin fermentation with S. pris(inaespiralis 12-55 is non-growth associated. Actinoplanes teichoniyceticus 97-5-74 for teicoplanin production had undergone mutation and screening with various methods. In this work, a valine hydroxamate resistant mutant strain was screened for the enhancement of valine production, which is the important precursor for teicoplanin TA22 synthesis. A VHr mutant strain A. eichomycehicus 98-1-227, which is high in total potency and TA22 titer, was obtained from the parent strain 97-5-74. The results of the shaking flask fermentation experiments suggested that teicoplanin fermentation with A. teichomyceticus 98-1-227 is also non-growth associated. Rifamycin B has been commercially produced by a strain of Amyco/atoposis mediterranei. In this work, an industrial applied strain XC 102 was used for further screening. A special mutation and screening procedure was adopted in this work to select a strain which could growth well on the medium with high concentration of VII 0.1% trp, 0.5% trp, 0.1% PHBA, and 0.1% propyl acid to alleviate the inhibition caused by both aromatic amino acid and PHBA in the metabolic pathway of rifamycin B, as well as to enhance the production of propyl acid which is the precursor of rifamycin B. By above methods, a strain A. niediterranei XC 9-25 was obtained. The rifamycin 13 productivity reaches l0000uIml, which is 1.38 times higher than that of the parent strain XC 102. Except for high productivity, strain XC 9-25 possesses the advantage of producing less mycclia, that is favorable in rifamycin recovery. According to the different characteristics of pristinamycin, teicoplanin, and rifamycin B fermentation, the scale-up of the fermentation processes for three antibiotics were studied, and the principles of scale-up were proposed. Pristinamycin fermentation needs high dissolved oxygen and is not very sensitive to shearing strength. Scale-up of pristinamycin fermentation from shaking flas

Full-text Catalog

Directory     3-7
Chinese Abstract     7-9
English summary     9-12
Preface     12-14
Chapter Literature Review     14-52
1.1 Introduction     14
1.2 the pristinamycin research progress     14-22
.2.1 Overview     14-16
1.2.2 pristinamycin biosynthesis     16-18
1.2.3 pristinamycin structural modification     18-22
1.3 teicoplanin research progress     22-24
Overview nbsp 1.3.1;   22-24
1. 3.2 teicoplanin biosynthesis     24
1.4 rifamycin progress     24-29
1.4.1 Overview     24-25 25-26
1.4.2 rifamycin biosynthesis    
1.4.3 the rifamycin genetics research     26-27
1.4.4 rifamycin biosynthetic metabolic regulation     the 27-29 1.4.5 rifamycin strain breeding
1.4.6 rifamycin fermentation and biotransformation     29
1.5 antibiotics bacteria the reasoning breeding     29-35
1.5. 1-resistant body and its structural analogues mutant screening     30
1.5.2 Resistance own metabolic screening of the final product and its structural analogues mutant     30-31
1.5.3 resistant carbon the catabolic repression mutant screening     31
1.5.4 resistant nitrogen catabolic repression mutant screening     31
1.5.5 resistant phosphate Mutants     31
1.5.6 induced the enzyme mutants Screening     31-32
1.5.7 antibiotic enzyme deletion mutant Filter     the the 32
1.5.8 morphology of Mutants     the mutant membrane permeability screening 32-33
1.5.9     33
1.5.10 primary metabolic pathways obstacles mutant screening   the   33-34
1.5.11 secondary metabolic pathways obstacles mutant Screening     34 < br /> 1.5.12 primary metabolic enzyme of high activity of mutant strains of     34
1.5.13 to adapt the the enzyme system regulation Mutants     34-35
1.6 antibiotic fermentation kinetics and dynamics model     the 35-42 1.6.1
antibiotics fermentation kinetics     35
1.6.2 the antibiotic fermentation kinetics model References nbsp     35-42
;   42-52
Chapter the pristinamycin fermentation conditions     nbsp 52-61
2.1 Introduction ;   52
2.2 Materials and methods     52-53
2.3 Results and discussion     53-59
2.3.1 pH of the original neomycin fermentation   the   53-54
2.3.2 inorganic nitrogen sources pristinamycin fermentation     54-55
2.3.3 organic nitrogen sources the pristinamycin fermentation     55-56
2.3.4 potassium dihydrogen phosphate, pristinamycin fermentation     56
2.3.5 carbon source pristinamycin fermentation     56-57
2.3.6 the organic alcohol pristinamycin fermented affect     57-58
2.3.7 soybean oil, pristinamycin fermentation     58
2.3.8 the Orthogonal Test pristinamycin fermentation medium     58-59
2.4 Summary     59-60
Reference     60-61
Chapter antibiotic producing bacteria reasoning breeding     61-94
3.1 Introduction     61
3.2 pristinamycin the producing bacteria reasoning breeding     61-71
3.2.1 Introduction     61-62
3.2.2 Materials and methods   the   62-64
3.2.3 Results and Discussion     64-69 glycine-resistant variants of screening     64 valine hydroxamic acid-resistant variants of screening     64-66
of kitasamycin resistant variants screening     66-67 2 - deoxy-D-glucose-resistant variants screening     67-68 strains 12 -55 the passages stability     68-69 Strains 12-55 of shake flask fermentation metabolism     69
3.2.4 Summary     69-71
3.3 teicoplanin produce the bacteria the reasoning breeding     71-78
3.3.1 foreword,     71
3. 3.2 Materials and methods     71-73
3.3.3 Results and discussion     73-78 strains, valine hydroxamate the acid sensitivity     73-74 valine hydroxamic acid-resistant variants Breeding     74 strains 98-1-227 genetic stability     74-75 strains 98-1-227 strains 97-1-74's production capacity and TA2-2 group component     75 the teicoplanin fermentation process of L-valine metabolic differences in     75-76
3.3.3. the 6 strains 98-1-227 flask fermentation metabolism     76-78
3.3.4 Summary     78
3.4 rifamycin producing bacteria reasoning election education     78-90
3.4.1 foreword,     78-79
3.4.2 Materials and methods nbsp the;   79-82
3. 4.3 Results and discussion     82-89 aromatic amino acid tolerant variant screening     83-85 pair the hydroxyl benzoic patience variant screening     85 propionate resistant variants screening     85-86 Strains XC -925 the passages stability     the the 86-87 strains, the XC-925 flask fermentation metabolism     87-89
3.4.4 Summary     89-90
3.5 Summary     the Reference 90
Chapter IV antibiotics the fermentation to enlarge     94 - 118
4.1 Introduction     94
4.2 pristinamycin the fermentation to enlarge     94-100
4.2.1 Materials and methods     95-96
4.2.2 Results and discussion     96-99 dissolved oxygen pristinamycin fermentation     96-97 < br /> shear force pristinamycin fermentation     the 2008 15L, fermenter batch fermentation     97-99
4.2.3 Summary     99-100
4.3 teicoplanin fermentation enlarge     100-105
4.3.1 Materials and methods     100-101
4.3.2 Results and discussion     101-104 dissolved oxygen fermentation teicoplanin     101 -102 shear force teicoplanin fermentation     102 15L fermenter batch fermentation     102 -104
4.3.4 Summary     104-105
4.4 rifamycin B fermentation enlarge     105-116
4.4.1 Materials and methods     105-107
4.4.2 Results and discussion     107-114 dissolved oxygen rifamycin B fermentation impact     107-108 shear force of rifamycin B fermentation     108 15L fermenter batch fermentation     108-109 15L fermenter fed - batch fermentation     109-110 fermenter enlarge     110-112 7m ~~ 3 fermenter fed-batch fermentation     112-114 60m ~~ 3 fermenter fed-batch fermentation     114
4.4.3 Summary     114-116
4.4 Summary     to References 116-117
    the antibiotic fermentation kinetics model 117-118
Chapter     118-128
5.1 Introduction     118
5.2 filamentous bacteria growth mechanism     118-120
5.3 antibiotic fermentation kinetic models     120-124
5.3.1 of mycelial the actual fermentation process classification     120
5.3.2 mycelial growth     120-121
5.3.3 mycelium, differentiation and inactivation     121
5.3.4 product formation     121-122
5.3.5 substrates. consumption     122-123
5.3.6 total model expression nbsp to;   123 - 124
5.4 of antibiotic fermentation process of fitting     124-126
5.5 Summary     the Reference 126
Chapter 6 Conclusions     128-130
Symbol Description     130-132
Acknowledgements     132

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