• Title/Summary/Keyword: fermentation inhibitor

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Effect of Organic Acids Derived from Black Liquor on Growth of Selected Escherichia coli MG 1655 (흑액 유래 유기산의 Escherichia coli MG 1655 성장에 미치는 영향 탐색)

  • Moon, Joon-Kwan;Um, Byung-Hwan
    • Journal of the Korean Wood Science and Technology
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    • v.42 no.6
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    • pp.758-767
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    • 2014
  • In this study, we selected an Escherichia coli strain (E. coli MG 1655) metabolizing arabinose derived from acid hydrolyzed black liquor as a carbon source and investigated effect of organic acids (acetic acid, formic acid, and lactic acid) presented in black liquor on growth of the E. coli MG 1655. We measured growth of E. coli MG 1655 under various concentration of each and combined three kinds of organic acids. The E. coli MG 1655 shows tolerance to acetic acid, lactic acid and formic acid at these concentrations ($1.0g/{\ell}$ acetic acid, $1.2g/{\ell}$ lactic acid and $0.8g/{\ell}$ formic acid, respectively), but displays some growth retardation over $1.5g/{\ell}$ acetic acid, lactic acid $2.0g/{\ell}$, and formic acid $1.2g/{\ell}$, respectively. In addition, formic acid was shown to be a critical factor affecting growth of the E. coli MG 1655 in the presence of three kinds of organic acids. These results indicate that the inhibitors should be removed at least $1.0g/{\ell}$ of acetic acid, $1.2g/{\ell}$ of lactic acid, $0.8g/{\ell}$ of formic acid for normal cell growth required for high yield fermentation. In addition, there is a need to construct recombinant strains that may be resistant to the same or higher organic acids concentration (> $1.2g/{\ell}$) in the growth.

Effects of Main Constituents of W/O Emulsion on Removal of Acetic Acid in a Simulated Hemicellulosic Hydrolysate (W/O 에멀젼의 주요 구성 성분들이 모사 헤미셀룰로오스 가수분해액에 있는 초산의 제거에 미치는 영향)

  • Lim, Sung Jin;Lee, Sang Cheol
    • Korean Chemical Engineering Research
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    • v.52 no.6
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    • pp.789-795
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    • 2014
  • Acetic acid is the most abundant and serious ethanol fermentation inhibitor in dilute-acid hemicellulosic hydrolysates. A mixture of xylose, acetic acid and sulfuric acid was chosen as a simulated hemicellulosic hydrolysate so as to find an optimal separation system to selectively remove acetic acid from the hydrolysates. In order to attain the purpose, emulsion liquid membrane was applied to removal of acetic acid from the simulated hemicellulosic hydrolysate. The effects of main constituents of water-in-oil (W/O) emulsion, such as amine extractant type, surfactant composition, additive type, and type and concentration of stripping agent, on extraction of acetic acid, xylose, and sulfuric acid in the simulated hemicellulosic hydrolysate were investigated. Under specific experimental conditions, degree of extraction of acetic acid was higher than 95% while loss of xylose was insignificant, which means that the current emulsion liquid membrane can be an economically feasible process.

Extracellular 5-Aminolevulinic Acid Production by Escherichia coli Containing the Rhodopseudomonas palustris KUGB306 hemA Gene

  • Choi, Han-Pil;Lee, Young-Mi;Yun, Cheol-Won;Sung, Ha-Chin
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1136-1140
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    • 2008
  • The Rhodopseudomonas palustris KUGB306 hemA gene codes for 5-aminolevulinic acid (ALA) synthase. This enzyme catalyzes the condensation of glycine and succinyl-CoA to yield ALA in the presence of the cofactor pyridoxal 5'-phosphate. The R. palustris KUGB306 hemA gene in the pGEX-KG vector system was transformed into Escherichia coli BL21. The effects of physiological factors on the extracellular production of ALA by the recombinant E. coli were studied. Terrific Broth (TB) medium resulted in significantly higher cell growth and ALA production than did Luria-Bertani (LB) medium. ALA production was significantly enhanced by the addition of succinate together with glycine in the medium. Maximal ALA production (2.5 g/l) was observed upon the addition of D-glucose as an ALA dehydratase inhibitor in the late-log culture phase. Based on the results obtained from the shake-flask cultures, fermentation was carried out using the recombinant E. coli in TB medium, with the initial addition of 90 mM glycine and 120 mM succinate, and the addition of 45 mM D-glucose in the late-log phase. The extracellular production of ALA was also influenced by the pH of the culture broth. We maintained a pH of 6.5 in the fermenter throughout the culture process, achieving the maximal levels of extracellular ALA production (5.15 g/l, 39.3 mM).

Enhancement of Clavulanic Acid Production by Expressing Regulatory Genes in gap Gene Deletion Mutant of Streptomyces clavuligerus NRRL3585

  • Jnawali, Hum Nath;Lee, Hei-Chan;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.146-152
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    • 2010
  • Streptomyces clavuligerus NRRL3585 produces a clinically important $\beta$-lactamase inhibitor, clavulanic acid (CA). In order to increase the production of CA, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene (gap) was deleted in S. clavuligerus NRRL3585 to overcome the limited glyceraldehyde-3-phosphate pool; the replicative and integrative expressions of ccaR (specific regulator of the CA biosynthetic operon) and claR (Lys-type transcriptional activator) genes were transformed together into a deletion mutant to improve clavulanic acid production. We constructed two recombinant plasmids to enhance the production of CA in the gap1 deletion mutant of S. clavuligerus NRRL3585: pHN11 was constructed for overexpression of ccaR-claR, whereas pHN12 was constructed for their chromosomal integration. Both pHN11 and pHN12 transformants enhanced the production of CA by 2.59-fold and 5.85-fold, respectively, compared with the gap1 deletion mutant. For further enhancement of CA, we fed the pHN11 and pHN12 transformants ornithine and glycerol. Compared with the gap1 deletion mutant, ornithine increased CA production by 3.24- and 6.51-fold in the pHN11 and pHN12 transformants, respectively, glycerol increased CA by 2.96- and 6.21-fold, respectively, and ornithine and glycerol together increased CA by 3.72- and 7.02-fold, respectively.

Isolation of Angiotensin I-Converting Enzyme Inhibitory Peptide from Chungkookjang (청국장으로부터 Angiotensin I 전환효소 저해 Peptide의 분리)

  • Matsui Toshiro;Yoo Hyung Jae;Hwang Jae Sung;Lee Dong Seok;Kim Han Bok
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.355-358
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    • 2004
  • Chunkookjang, Korean traditional fermented soybean food emerges as a functional food to improve intestinal function and blood circulation. During Chungkookjang fermentation, microorganisms, enzymes, and diverse bioactive compounds increase sharply. Chungkookjang contains diverse oligo-peptides. Formation of peptides was confirmed by SDS-PAGE. Solube fermented soybean in our sample contained Tyr, Gln-Lys, Trp, Gln, and Lys-Pro as major components. Lys-Pro (0.083 mg/100 g sample) was purified by HPLC analysis. Angiotensin I­converting enzyme (ACE) causes hypertension by converting angiotensin I to angiotension II. ACE inhibitory activity of Lys-Pro was determined to be $IC_{50}=32.1\;{\mu}M.$ Whether or not eating Chungkookjang can lower blood pressure was also determined. Sistolic blood pressure dropped by 15 mmHg, and diastolic blood pressure by 8 mmHg 2 hr after a single administration of 20 g of fermented soybean. Chungkookjang might be helpful in improving blood circulation since it has ACE inhibitor and antihypertenisve effect.

Isolation of Mutant Yeast Strains having Resistance to 1-ethyl-3-methylimidazolium Acetate through a Directed Evolutionary Approach (유도적 돌연변이 유발 방법을 통한 1-ethyl-3-methylimidazolium acetate에 대해 내성을 갖는 돌연변이 효모 선별)

  • Lee, Yoo-Jin;Kwon, Deok-Ho;Park, Jae-Bum;Ha, Suk-Jin
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.51-56
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    • 2017
  • Cellulosic biomass is a renewable source for biofuel production from non-edible biomass. An optimized pretreatment process is required for the efficient utilization of cellulosic biomass. Among various pretreatment processes, the use of ionic liquids has been reported recently. However, the residual ionic liquid after pretreatment acts as an inhibitor of microbial fermentation. Recently, we isolated mutant Saccharomyces cerevisiae strains resistant to the ionic liquid 1-ethyl-3-methylimidazolium acetate ([EMIM][Ac]) by using a directed evolutionary approach. When 3% [EMIM][Ac] was added to a medium containing 80 g/l of glucose, mutants D452-B2 and D452-S3 produced 35.6 g/l and 36.3 g/l of ethanol, respectively, for 18 h while the parental strain (S. cerevisiae D452-2) produced 1.3 g/l of ethanol. Thus, these mutant S. cerevisiae strains might prove advantageous when ionic liquids are used for biofuel production from cellulosic biomass.

The Suppressive Effect of Pueraria lobata Root Extract and Its Biotransformed Preparation against Skin Wrinkle Formation

  • Koo, Hyun Jung;Lee, SungRyul;Kang, Se Chan;Kwon, Jung Eun;Lee, Da Eun;Choung, Eui-Su;Lee, Jong-Sub;Lee, Jin Woo;Park, Yuna;Sim, Dong Soo;Sohn, Eun-Hwa
    • Korean Journal of Plant Resources
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    • v.30 no.3
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    • pp.272-279
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    • 2017
  • EP was obtained through 20% ethanol extraction of Pueraria lobata root, and the fermented form of EP, FEP, was prepared from the EP after incubating with Lactobacillus rhamnosus vitaP1. There was no significant toxicity by EP and FEP up to $1000{\mu}g/ml$ in NIH-3T3, HaCaT, and B16F10 cells. In addition to antioxidant potentials of EP and FEP determined by DPPH and ABST assays, we confirmed increase of procollagen type I and elastin synthesis by supplementation of the EP and FEP at the concentration of $50{\mu}g/ml$ using ELISA kits. The protein expression levels of matrix metalloprotease (MMP)-1, -3, and -9, those are involved in the degradation of collagen or other skin matrix proteins, were remarkably suppressed while their inhibitory protein metallopeptidase inhibitor 1 (TIMP-1) was greatly up-regulated by supplementation of the EP and FEP at a concentration of $50{\mu}g/ml$. Taken together, both EP and FEP supplementation could be involved in the suppression of the skin wrinkle formation through inhibiting degradation of collagen and stimulating the synthesis of collagen and elastin. The results showed that the anti-wrinkle potential of the EP and FEP will be a promising candidate for developing cosmeceutical compounds or products.

Mitophagy Improves Ethanol Tolerance in Yeast: Regulation by Mitochondrial Reactive Oxygen Species in Saccharomyces cerevisiae

  • Jing, Hongjuan;Liu, Huanhuan;Lu, Zhang;Cui, liuqing;Tan, Xiaorong
    • Journal of Microbiology and Biotechnology
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    • v.30 no.12
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    • pp.1876-1884
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    • 2020
  • Ethanol often accumulates during the process of wine fermentation, and mitophagy has critical role in ethanol output. However, the relationship between mitophagy and ethanol stress is still unclear. In this study, the expression of ATG11 and ATG32 genes exposed to ethanol stress was accessed by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). The result indicated that ethanol stress induced expression of the ATG11 and ATG32 genes. The colony sizes and the alcohol yield of atg11 and atg32 were also smaller and lower than those of wild type strain under ethanol whereas the mortality of mutants is higher. Furthermore, compared with wild type, the membrane integrity and the mitochondrial membrane potential of atg11 and atg32 exhibited greater damage following ethanol stress. In addition, a greater proportion of mutant cells were arrested at the G1/G0 cell cycle. There was more aggregation of peroxide hydrogen (H2O2) and superoxide anion (O2•-) in mutants. These changes in H2O2 and O2•- in yeasts were altered by reductants or inhibitors of scavenging enzyme by means of regulating the expression of ATG11 and ATG32 genes. Inhibitors of the mitochondrial electron transport chain (mtETC) also increased production of H2O2 and O2•- by enhancing expression of the ATG11 and ATG32 genes. Further results showed that activator or inhibitor of autophagy also activated or inhibited mitophagy by altering production of H2O2 and O2•. Therefore, ethanol stress induces mitophagy which improves yeast the tolerance to ethanol and the level of mitophagy during ethanol stress is regulated by ROS derived from mtETC.

Mechanisms of Insulinotropic Effect of YHB-2017 [Genistein] Isolated from fermentation Broths of Streptomyces sp. (방선균에서 유래한 YHB-2017 [Genistein]의 인슐린 분비 촉진 작용 기전)

  • Kwag, Won-Jae;Park, You-Hoi;Park, Jun-Chul;Lee, Byung-Kyu;Kang, Yup;Choe, Tae-Boo
    • KSBB Journal
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    • v.21 no.6 s.101
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    • pp.466-473
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    • 2006
  • Impaired insulin secretion from pancreatic beta-cells in response to glucose is an important feature in the pathology of non-insulin-dependent diabetes mellitus (NIDDM). In the course of screening for useful insulin secretagogues, we have isolated and identified YHB-2017 (Genistein) as a insulin secretion potentiator from fermentation broths of our in-house microbial library. The insulinotropic activity of YHB-2017 in isolated rat pancreatic islets was exerted only at high concentration of glucose (8.3-16 mM) but not at low concentration of glucose (3.3-5.5 mM). Also, in perifusion study with isolated rat pancreatic islets, YHB-2017 stimulated insulin secretion in a time-dependent manner when YHB-2017 was added to KRB buffer containing 16 mM glucose. In the presence of $200\;{\mu}M$ diazoxide and 35 mM KCI, which stimulates maximum $Ca^{2+}$ influx independently of KATP channel, YHB-2017 enhanced KATP channel-independent insulin secretion at high concentration glucose (16 mM). To elucidate the mechanisms of the glucose-dependent potentiation effect of YHB-2017, pharmacologic inhibitors for protein kinase A, protein kinase C and calcium/calmodulin kinase II were pre-treated and then the potentiation effect of YHB-2017 on insulin secretion was investigated. Pre-treatment of H89 as a PKA inhibitor had a significant inhibitory effect on YHB-2017-induced potentiation effect. Furthermore, western immunoblotting analyses revealed that YHB-2017 increased phosphorylation of PKA substrates and cAMP response element-binding protein (CREB) under high concentration of glucose. These results demonstrated that the insulinotropic effect of YHB-2017 is mediated through PKA signal pathway and activated amplifying $K_{ATP}$ channel-independent insulin secretion pathway.

Effects of Physico-chemical and Microbiological Inhibitors for Odour gas Evolution in the Fermentation of Liverstock Feces (가축(家畜) 분뇨(糞尿) 발효시(醱酵時) 악취(惡臭)가스 생성억제제(生成抑制劑) 시용(施用) 효과에 관(關)한 연구(硏究))

  • Yun, Sei-Young;Lee, Sang-Kyu
    • Korean Journal of Soil Science and Fertilizer
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    • v.25 no.1
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    • pp.62-69
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    • 1992
  • A series of laboratory experiment was conducted to find out the effects of physico-chemical, microbiological and commercial inhibitors on the odorous gas evolution in the fermentation of livestock feces. The results obtained were summarized as follows. 1. The rate and amount of evolution of gas were the highest at 7 days after incubation, thereafter gradually decreased until 24 days after incubation. 2. The rate and amount of gases were evolved in order of $CO_2>N_2O>CH_4>NH_3>N_2S$, respectively. 3. The highest amount of methane gas was evolved from the poultry feces, those of carbon dioxide and nitrous oxide were evolved from the pig feces, and that of hydrogen sulfide was dominantly evolved from the cattle feces. 4. Negative correlation were obtained between the total amounts of $NH_3$ and $CH_4$, $CO_2$ and $CH_4$, $N_2O$ and $CH_4$, $N_2O$ and $CH_4$, while positive correlations were obtained between the amounts of $CO_2$ and $N_2O$, $CO_2$ and $NH_3$, and $NH_3$ and $N_2O$, respectively. 5. There was no significantly inhibiting effect obtained that the application of commercial gas inhibitor as VK 88. On the other hand there was significantly inhibiting effect obtained when application of fertile paddy soil and photosynthetic bacteria to the fermentation of livestock feces.

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