• Title/Summary/Keyword: Recombinant fermentation

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Change in Proteomic Profiles of Genetically Modified 1,3-Propanediol-Producing Recombinant E. coli

  • Jin, Li-Hua;Lee, Jung-Heon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1439-1444
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    • 2008
  • The recombinant E. coli $\Delta$6 mutant (galR, glpK, gldA, IdhA, lacI, tpiA) was used to produce 1,3-propanediol (PD) from glucose. The 1,3-PD production increased with feedback control of the glucose concentration using fed-batch fermentation. The maximum 1,3-PD concentration produced was 43 g/l after 60 h of fermentation. Glycerol production was minimized when controlling the glucose concentration at less than 1 g/l. The expression levels of seven enzymes related to the 1,3-PD production metabolism were compared during the cell growth phase and 1,3-PD production phase, and their expression levels all increased during 1,3-PD production, with the exception of alcohol dehydrogenase.

Hybridization에 의한 반수체 재조합 효모균주의 전분 발효능 증진

  • Park, Sun-Young;Kim, Keun;Lee, Chang-Hoo
    • Microbiology and Biotechnology Letters
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    • v.24 no.6
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    • pp.726-732
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    • 1996
  • To improve the fermentation characteristics(such as starch-degradability, ethanol tolerance, sugar and high-temperature tolerance) of recombinant haploid yeast Saccharomyces diastaticus K114, hybridization technique was used. The hybridization partner was S. diastaticus 1177 which had good glucoamylase activity and fermentabi- lity. The best hybrid HH64 showed improved ethanol tolerance, sugar and high-temperature tolerance. Especia- lly, the starch-fermentability was significantly improved, since the hybrid produced 1.60% (w/v) ethanol from 4% (w/v) starch, while the recombinant haploid K114 produced 1.30% (w/v) ethanol. The optimum temperature and pH for the starch-fermentation by the hybrid HH64 was 30$\circ$C and 5, respectively. The hybrid yeast HH64 produced 7.5% (w/v) ethanol directly from 20% (w/v) starch.

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Increased Production of Recombinant Protein by Escherichia coli Deficient in Acetic Acid Formation

  • Koo, Tae-Young;Park, Tai-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.789-793
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    • 1999
  • The effect of acetic acid formation deficiency on recombinant E. coli fermentation was investigated using a mutant strain deficient in acetic acid formation. A mutant strain which does not grow under anaerobic conditions was isolated. The acetic acid production in this strain was negligible in aerobic batch fermentation. The cloned-gene expression in the mutant strain was higher than the wild-type strain. Fed-batch fermentations with controlled specific growth rates were carried out in order to compare the cloned-gene expression between the wild-type and the mutant strains. The expression decreased along with the specific growth rate in both strains. The cloned-gene expression in the mutant strain was 60% higher than in the wild-type strain at the same specific growth rate.

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Polyaromatic Resin HP-20 Induced Accumulation of Intermediate Azaphilones in Monascus purpureus 𝚫mppC and 𝚫mpp7 Strains

  • Lim, Yoon Ji;Lee, Doh Won;Choi, Jeong Ju;Park, Si-Hyung;Kwon, Hyung-Jin
    • Journal of Microbiology and Biotechnology
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    • v.29 no.6
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    • pp.897-904
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    • 2019
  • Monascus purpureus recombinant mppC and mpp7 knockout strains were subjected to extractive fermentation in the context of azaphilone pigment production. Inclusion of Diaion HP-20 resin resulted in the selective production of unreduced azaphilone congeners, in addition to the early intermediate FK17-P2a, from ${\Delta}mppC$ and ${\Delta}mpp7$ strains that would otherwise mainly produce reduced congeners. Structural determination of two novel unreduced azaphilones from the ${\Delta}mpp7$ strain was accomplished. The unreduced azaphilone compound was converted into the cognate reduced congener in recombinant M. purpureus strains, demonstrating its intermediate role in azaphilone biosynthesis. This study demonstrates the possibility that extractive fermentation with Diaion HP-20 resin can be used to obtain cryptic azaphilone metabolites.

Construction and Analysis of Food-Grade Lactobacillus kefiranofaciens β-Galactosidase Overexpression System

  • He, Xi;Luan, MingJian;Han, Ning;Wang, Ting;Zhao, Xiangzhong;Yao, Yanyan
    • Journal of Microbiology and Biotechnology
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    • v.31 no.4
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    • pp.550-558
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    • 2021
  • Lactobacillus kefiranofaciens contains two types of β-galactosidase, LacLM and LacZ, belonging to different glycoside hydrolase families. The difference in function between them has been unclear so far for practical application. In this study, LacLM and LacZ from L. kefiranofaciens ATCC51647 were cloned into constitutive lactobacillal expression vector pMG36e, respectively. Furtherly, pMG36n-lacs was constructed from pMG36e-lacs by replacing erythromycin with nisin as selective marker for food-grade expressing systems in Lactobacillus plantarum WCFS1, designated recombinant LacLM and LacZ respectively. The results from hydrolysis of o-nitrophenyl-β-galactopyranoside (ONPG) showed that the β-galactosidases activity of the recombinant LacLM and LacZ was 1460% and 670% higher than that of the original L. kefiranofaciens. Moreover, the lactose hydrolytic activity of recombinant LacLM was higher than that of LacZ in milk. Nevertheless, compare to LacZ, in 25% lactose solution the galacto-oligosaccharides (GOS) production of recombinant LacLM was lower. Therefore, two β-galactopyranosides could play different roles in carbohydrate metabolism of L. kefiranofaciens. In addition, the maximal growth rate of two recombinant strains were evaluated with different temperature level and nisin concentration in fermentation assay for practical purpose. The results displayed that 37℃ and 20-40 U/ml nisin were the optimal fermentation conditions for the growth of recombinant β-galactosidase strains. Altogether the food-grade Expression system of recombinant β-galactosidase was feasible for applications in the food and dairy industry.

Effect of Acetic Acid Formation and Specific Growth Rate on Productivity of Recombinant Escherichia coli Fed-Batch Fermentation (초산 생성 및 비성장속도가 재조합 대장균 유가식 발효의 생산성에 미치는 영향)

  • 구태영;박태현
    • KSBB Journal
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    • v.10 no.4
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    • pp.455-460
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    • 1995
  • Specific growth rate was controlled for the repression of acetic acid formation in the fed-batch fermentation of recombinant Escherichia coli. With controlled specific growth rate, we studied the effect of the specific growth rate on cell growth, glucose consumption, acetic acid formation, and the expression of recombinant protein (${\beta}$-lactamase). High specific growth rate caused the accumulation of glucose and acetic acid, and lowered the production of recombinant protein. However, the addition of methionine recovered the gene expression by alleviating the negative effect of acetic acid at high specific growth rate.

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Fermentation Process Development of Recombinant Hansenula polymorpha for Gamma-Linolenic Acid Production

  • Khongto, B.;Laoteng, K.;Tongta, A.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1555-1562
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    • 2010
  • Development of the strain and the fermentation process of Hansenula polymorpha was implemented for the production of ${\gamma}$-linolenic acid ($GLA,\;C18:3{\Delta}^{6,9,12}$), an n-6 polyunsaturated fatty acid (PUFA) that has been reported to possess a number of health benefits. The mutated ${\Delta}^6$-desaturase (S213A) gene of Mucor rouxii was expressed in H. polymorpha under the control of the methanol oxidase (MOX) promoter. Without the utilization of methanol, a high-cell-density culture of the yeast recombinant carrying the ${\Delta}^6$-desaturase gene was then achieved by fed-batch fermentation under glycerol-limited conditions. As a result, high levels of the ${\Delta}^6$-desaturated products, octadecadienoic acid ($C18:2{\Delta}^{6,9}$), GLA, and stearidonic acid ($C18:4{\Delta}^{6,9,12,15}$), were accumulated under the derepression conditions. The GLA production was also optimized by adjusting the specific growth rate. The results show that the specific growth rate affected both the lipid content and the fatty acid composition of the GLA-producing recombinant. Among the various specific growth rates tested, the highest GLA concentration of 697 mg/l was obtained in the culture with a specific growth rate of 0.08 /h. Interestingly, the fatty acid profile of the yeast recombinant bearing the Mucor ${\Delta}^6$-desaturase gene was similar to that of blackcurrant oil, with both containing similar proportions of n-3 and n-6 essential fatty acids.

Effect of Oxygen Supply on the Production of Interferon ${\alpha}$-1 by Recombinant Escherichia coli in Fed-batch Fermentation (유가식 배양에서 재조합 대장균으로부터 Interferon ${\alpha}$-1 생산에 산소 공급이 미치는 영향)

  • Yi, Jong-Ghil;Moon, Seok-Young;Kim, Young-Jun;Shin, Chul-Soo;Koo, Yoon-Mo
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.226-230
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    • 2007
  • In order to achieve high-level expression of interferon-${\alpha}1$ (IFN-${\alpha}1$) during fed-batch fermentation of recombinant E. coli, effects of oxygen supply and induction temperature on the expression of recombinant proteins were evaluated. Supplementation of oxygen and its transfer into cells is one of the most important parameters involved in the design and operation of mixing-sparging equipment for bioreactors. Generally, higher oxygen supply stimulates cell growth of aerobic microorganism and consequently the amount of products is increased. In this study, the optimum aeration strategy for the higher production of IFN-${\alpha}1$ during fed-batch fermentation of recombinant E. coli was surveyed. The growth of the cells was also monitored with four different concentrations of dissolved oxygen (DO; limiting, 20%, 35%, 50%) conditions. The DO was controlled by varying aeration rates of air and pure oxygen. Oxygen uptake rate (OUR) and specific oxygen uptake rate (SOUR) were evaluated and compared for the enhanced growth and induction of the cells and IFN-${\alpha}1$, respectively. We confirmed that increased DO by additional oxygen supply, up to 35%, can improve the production of IFN-${\alpha}1$ during the fermentation.

A Fermentation Strategy for Anti-MUC1 C595 Diabody Expression in Recombinant Escherichia Coli

  • Lan, John Chi-Wei;Ling, Tau Chuan;Hamilton, Grant;Lyddiatt, Andrew
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.425-431
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    • 2006
  • The development of fermentation conditions for the production of C595 diabody fragment (dbFv) in E. coli HB2151 clone has been explored. Investigations were carried out to study the effect of carbon supplements over the expression period, the comparison of C595 dbfv production in synthetic and complex media, the influence of acetic acid upon antibody production, and comparison of one-stage and two-stage processes operated at batch or fed-batch modes in bioreactor. Yeast extract supplied during expression yielded more antibody fragment than any other carbon supply. The synthetic medium presented higher specific productivity (0.066 mg dbFv $g^{-1}$ dry cell weight) when compared to the complex medium (0.044 mg dbFv $g^{-1}$ DCW). The comparison of fermentation strategies demonstrated that (1) one-stage fed-batch fermentation performed higher C595 dbFv production than that operated in batch mode which was significantly affected by acetate concentration; (2) a two-stage batch operation could enhance C595 dbFv production. It was found that a concentration of 12.3 mg $L^{-1}$ broth of C595 dbFv and a cell concentration of 10.8g $L^{-1}$ broth were achieved at the end of two-stage operation in 5-L fermentation.