• Title/Summary/Keyword: sole carbon source

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Isolation of a Pseudomonas sp. Strain Exhibiting Unusual Behavior of Poly(3-hydroxyalkanoates) Biosynthesis and Characterization of Synthesized Polyesters

  • Chung, Chung-Wook;Kim, Yoon-Seok;Kim, Young-Baek;Bae, Kyung-Sook;Rhee, Young-Ha
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.847-853
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    • 1999
  • A Pseudomonas sp. strain that is capable of utilizing dicarboxylic acids as a sole carbon source was isolated from activated sludge by using the enrichment culture technique. This organism accumulated polyhydroxyalkanoates (PHAs) with an unusual pattern of monomer units that depends on the carbon sources used. Polyhydroxybutyrate (PHB) homopolyester was synthesized from glucose or small $C_{-even}$ alkanoic acids, such as butyric acid and hexanoic acid. Accumulation of PHB homopolyester was also observed in the cells grown on $C_{-odd}$ dicarboxylic acids, such as heptanedioic acid and nonanedioic acid as the sole carbon sources. In contrast, a copolyester consisting of 6 mol% 3-hydroxybutyrate (3HB) and 94 mol% 3-hydroxyvalerate (3HV) was produced with a PHA content of as much as 36% of the cellular dry matter. This strain produced PHAs consisting both of the short-chain-length (SCL) and the medium-chain-length (MCL) 3-hydroxyacid units when heptanoic acid to undecanoic acid were fed as the sole carbon sources. Most interestingly, polyester consisting of significant amount of relevant fractions, 3HB, 3HV, and 3-hydroxyheptanoate (3HHp), was accumulated from heptanoic acid. According to solvent fractionation experiments, the polymer produced from heptanoic acid was a blend of poly(3HHp) and of a copolyester of 3HB, 3HV, and 3HHp units. The hexane soluble fractions contained only 3HHp units while the hexane-insoluble fractions contained 3HB and 3HV units with a small amount of 3HHp unit. The copolyester was an elastomer with unusual mechanical properties. The maximum elongation ratio of the copolyester was 460% with an ultimate strength of 10 MPa, which was very different from those of poly(3HB-co-3HV) copolyesters having similar compositions produced from other microorganisms.

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Site-specific Disruption of Glyoxylate Bypass and Its Effect in Lysine-producing Corynebacterium lactofermentum Strain

  • Kim, Youn-Hee;Lee, Heung-Shick
    • Journal of Microbiology and Biotechnology
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    • v.6 no.5
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    • pp.315-320
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    • 1996
  • The role of glyoxylate bypass in a lysine-producing Corynebacterium lactofermentum strain was analyzed. Unlike the wild type, the strain expressed enzymes of glyoxylate bypass during growth in the fermentation broth containing glucose as the carbon source. To evaluate the importance of glyoxylate bypass in the strain, we disrupted chromosomal aceA by using a cloned fragment of the gene. Site-specific disruption of aceA which codes for the isocitrate lyase, the first enzyme of the bypass, was confirmed by Southern blot analysis. The aceA mutant strain completely lost isocitrate lyase activity and ability to grow in a minimal medium containing acetate as the sole carbon source. The mutant strain was similar to its parental strain in growth characteristics and produced comparable amounts of lysine in shake flasks containing glucose as the carbon source. The amount of oxaloacetate accumulated in the fermentation medium was similar for both strains, suggesting that expression of glyoxylate bypass does not necessarily lead to the increase in intracellular oxaloacetate. These data clearly demonstrate that glyoxylate bypass does not function as one of the routes of carbon supply for lysine production in the strain. It appears that the leakiness of the glyoxylate bypass in the strain might be the result of a secondary mutation which arose during previous strain development by random mutagenesis.

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Roles of Glucose and Acetate as Carbon Sources in L-Histidine Production with Brevibacterium flavum FERM1564 Revealed by Metabolic Flux Analysis

  • Shioya, Suteaki;Shimizu, Hiroshi;Shimizu, Nobuyuki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.3
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    • pp.171-177
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    • 2002
  • The metabolic flux pattern for L-histidine production was analyzed when glucose and/or acetate were used as carbon sources. Total L-histidine production was enhanced when mixed substrate (glucose and acetate) was used, compared wish that when either glucose or acetate was used as the sole carbon source. Theoretical maximum carbon fluxes through the main pathways for L-histldine production, cell growth, and ATP consumption for cell maintenance were obtained by the linear programming (LP) method. By comparison of the theoretical maximum carbon fluxes tilth actual ones, it was found that a large amount of glucose was actually used for maintenance of cell viability. On the other hand, acetate was used for cell growth. After depletion of acetate in the mixed substrate culture, the flux for glucose to L-histldine synthesis was markedly enhanced. A strategy for effective L-histidine production using both carbon sources was proposed.

S5 Lipase : An Organic Solvent Tolerant Enzyme

  • Zaliha Raja Noor;Rahman Raja Abdul;Baharum Syarul Nataqain;Salleh Abu Bakar;Basri Mahiran
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.583-590
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    • 2006
  • In this study, an organic solvent tolerant bacterial strain was isolated. This strain was identified as Pseudomonas sp. strain S5, and was shown to degrade BTEX (Benzene, Toluene, Ethyl-Benzene, and Xylene). Strain S5 generates an organic solvent-tolerant lipase in the late logarithmic phase of growth. Maximum lipase production was exhibited when peptone was utilized as the sole nitrogen source. Addition of any of the selected carbon sources to the medium resulted in a significant reduction of enzyme production. Lower lipase generation was noted when an inorganic nitrogen source was used as the sole nitrogen source. This bacterium hydrolyzed all tested triglycerides and the highest levels of pro-duction were observed when olive oil was used as a natural triglyceride. Basal medium containing Tween 60 enhanced lipase production to the most significant degree. The absence of magnesium ions ($Mg^{2+}$) in the basal medium was also shown to stimulate lipase production. Meanwhile, an alkaline earth metal ion, $Na^+$, was found to stimulate the production of S5 lipase.

Carbon Source-Dependent Regulation of the Schizosaccharomyces pombe pbh1 Gene

  • Kim, Su-Jung;Cho, Nam-Chul;Ryu, In-Wang;Kim, Kyung-Hoon;Park, Eun-Hee;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.689-693
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    • 2006
  • Pbh1, from the fission yeast Schizosaccharomyces pombe, is a baculoviral inhibitor of apoptosis (IAP) repeat (BIR) domain-containing protein. Its unique encoding gene was previously found to be regulated by nitric oxide and nitrogen starvation. In the current work, the Pbh1-lacZ fusion gene was used to elucidate the transcriptional regulation of the pbh1 gene under various carbon sources. When fermentable carbon sources, such as glucose (at a low concentration of 0.2 %), sucrose (2.0 %) and lactose (2.0 %), were the sole carbon source, the synthesis of $\beta$-galactosidase from the Pbh1-lacZ fusion gene was reasonably enhanced. However, the induction by these fermentable carbon sources was abolished in the Pap1-negative S. pombe cells, implying that this type of induction of the pbh1 gene is mediated by Pap1. Ethanol (2.0%), a nonfermentable carbon source, was also able to enhance the synthesis of $\beta$-galactosidase from the fusion gene in wild-type cells but not in Pap1-negative cells. The results indicate that the S. pombe pbh1 gene is up-regulated under metabolic oxidative stress in a Pap1-dependent manner.

Enhancement of $\beta$-D-Glucans Production by Agaricus blazei Murill by Nitrogen Supplementation

  • NA JEONG-GEOL;KIM HYUN-HAN;CHUN GIE-TAEK;CHANG YONG KEUN;LEE SANG JONG;CHUNG YEON HO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1388-1391
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    • 2005
  • Temporal changes of cell growth pattern and intracellular content of $\beta$-D-glucans were investigated with off-gas data in Agaricus blazei culture where glucose was intermittently fed. It was observed that the time point of carbon source depletion coincided with the point of sudden drop in the carbon dioxide evolution rate (CER), and that the sole supplementation of glucose was not enough to maintain active cell growth and glucan content. On the other hand, when yeast extract, a typical nitrogen source, was supplemented together with glucose when the CER suddenly dropped because of carbon source depletion, an active cell growth could be maintained until the end of the culture and the glucan content did not decrease with culture time, significantly enhancing glucan productivity.

Degradation of Indole by Acinetobacter sp. KL-9 with Production of Indigo (Acinetobacter sp. KL-9에의한 indole 분해 및 Indigo 생성의 특성)

  • Kim, Jin-Wan;Kim, Jin-Kyoo;Lee, Soo-O;Lee, Kyoung
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.43-49
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    • 2001
  • Indole and its derivatives form a class of toxic recalcitrant environmental pollutants, Abacte-rium, strain KL-9 was isolated from soil with indole as a sole source of carbon and nitrogen. KL-9 was identified as Acinetobacter sp. on the basis of 16 S rRNA gene sequence, fatty acid and quinone compositions. This identification was also confirmed by the ability of carbon source utilization and other biochemical tests. The growth of Acinetobacter sp. KL-9 was fastest with 0.3mg/ml of indole as was inhibited by higher than 0.5mg/ml of indole in the medium, KL-9 with indole also produced indigo. The formation of indigo was stimulated inthe presence of glucose, which is not a growth-suppoting carbon source for KL-9. Additional biotransformation evidence showed that anthranilate is an intermediate for the degradation of indole KL-9.

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Regulation of $\beta$-Xylosidase (XylA) Synthesis in Bacillus stearothermophilus

  • Cho, Ssang-Goo;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.14-20
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    • 1998
  • Syntheses of the B. stearothermophilus xylanolytic enzymes such as xylanases, ${\beta}$-xylosidases, ${\alpha}$-arabinofurano-sidases, and esterases, were observed to be regulated by the carbon source present in the culture media. Xylan induced synthesis of ${\beta}$-xylosidase at the highest level while xylose gave about 30% of the ${\beta}$-xylosidase activity induced by xylan. The lowest syntheses of the xylanolytic enzymes above mentioned were detected in the basal medium containing glucose as a sole carbon source. When a mixture of xylan and glucose was used as a carbon source, we could observe glucose repression of xylanase (about 70-fold) and ${\beta}$-xylosidase (about 40-fold) syntheses. Whereas, the level of the glucose repression of the expression of the xylA gene encoding the major ${\beta}$-xylosidase of B. stearothermophilus was assessed to be about l0-fold when the relative amounts of the xylA transcript were determined. From the sequence of the xylA gene, we could find two CRE-like sequences (CRE-l: nucleotides +124 to +136 and CRE-2:+247 to +259) within the reading frame of the xylA gene, either or both of which were suspected to be involved in catabolite repression of the xylA gene.

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Dynamic Gene Expression Profiling of Escherichia coli in Carbon Source Transition from Glucose to Acetate

  • Oh Min-Kyu;Cha Mee-Jeong;Lee Sun-Gu;Rohlin Lars;Liao James C.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.543-549
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    • 2006
  • DNA microarray was used to study the transcription profiling of Escherichia coli adapting to acetate as a sole carbon source. Bacteria grown in glucose minimal media were used as a reference. The dynamic expression levels of 3,497 genes were monitored at seven time points during this adaptation. Among the central metabolic genes, the glycolytic and glucose phosphotransferase genes were repressed as the bacteria entered stationary phase, whereas the glyoxylate pathway, TCA cycle, and gluconeogenic genes were induced. Distinct induction or repression patterns were recognized among different pathway genes. For example, the repression of glycolytic genes and the induction of gluconeogenic ones started immediately after glucose was depleted. On the other hand, the regulation of the pentose phosphate pathway genes and glyoxylate genes gradually responded to the glucose depletion or was more related to growth in acetate. When the whole genome was considered, many of the CRP, FadR, and Cra regulons were immediately responsive to the glucose depletion, whereas the $\sigma^s$, Lrp, and IHF regulons were gradually responsive to the glucose depletion. The expression profiling also provided differential regulations between isoenzymes; for example, malic enzymes A (sfcA) and B (maeB). The expression profiles of three genes were confirmed with RT-PCR.

Overexpression of Mutant Galactose Permease (ScGal2_N376F) Effective for Utilization of Glucose/Xylose or Glucose/Galactose Mixture by Engineered Kluyveromyces marxianus

  • Kwon, Deok-Ho;Kim, Saet-Byeol;Park, Jae-Bum;Ha, Suk-Jin
    • Journal of Microbiology and Biotechnology
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    • v.30 no.12
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    • pp.1944-1949
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    • 2020
  • Mutant sugar transporter ScGAL2-N376F was overexpressed in Kluyveromyces marxianus for efficient utilization of xylose, which is one of the main components of cellulosic biomass. K. marxianus ScGal2_N376F, the ScGAL2-N376F-overexpressing strain, exhibited 47.04 g/l of xylose consumption and 26.55 g/l of xylitol production, as compared to the parental strain (24.68 g/l and 7.03 g/l, respectively) when xylose was used as the sole carbon source. When a mixture of glucose and xylose was used as the carbon source, xylose consumption and xylitol production rates were improved by 195% and 360%, respectively, by K. marxianus ScGal2_N376F. Moreover, the glucose consumption rate was improved by 27% as compared to that in the parental strain. Overexpression of both wild-type ScGAL2 and mutant ScGAL2-N376F showed 48% and 52% enhanced sugar consumption and ethanol production rates, respectively, when a mixture of glucose and galactose was used as the carbon source, which is the main component of marine biomass. As shown in this study, ScGAL2-N376F overexpression can be applied for the efficient production of biofuels or biochemicals from cellulosic or marine biomass.