• Journal of Microbiology and Biotechnology
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• 제18권3호
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• pp.539-544
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• 2008

A mutant of Arthrospira platensis PCC 9108, strain M9108, obtained by mutagenesis with UV treatment, was able to mixotrophically grow in an SOT medium containing 40 g of glucose/l. The biomass and specific growth rate of strain M9108 (4.10 g/l and 0.70/d) were 1.9-fold and 1.4-fold higher, respectively, than those of the wild type (2.21 g/l and 0.58/d) under mixotrophic culture condition. In addition, when compared with the wild type, the content of ${\gamma}$-linolenic acid (GLA) in the mutant was increased when glucose concentration was increased. Compared with the wild type, the GLA content of the mutant was 2-fold higher in autotrophic culture and about 3-fold higher in mixotrophic culture. Thus, the mutant appears to possess more efficient facility to assimilate and metabolize glucose and to produce more GLA than its wild-type strain.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.189-192
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• 2000

광합성 홍색세균의 light harvesting complex II (LHC II)발현 유전자가 제거된 돌연변이종을 halogen 램프 하에서 거리에 따라 광도를 달리하며 배양하여 wild type과 생산성을 비교한 결과 낮은 광도(34 ${\mu}E/m^2/s$)에서는 wild type가 높은 광도 (376, 118 ${\mu}E/m^2/s$)에서는 mutant가 높은 세포농도를 나타내었다. 특히 118 ${\mu}E/m^2/s$에서 LHC $II^-$ mutant가 56% 높은 세포생산량을 보였다. 이는 세포내 pigment양의 감소로 mutual shading effect가 감소하였기 때문으로 판단되었다.

• Parasites, Hosts and Diseases
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• 제54권1호
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• pp.55-60
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• 2016

The aim of the present study is to determine the characteristics of genotype and phenotype of Echinococcus granulosus derived from wild sheep and to compare them with the strains of E. granulosus sensu stricto (sheep-dog) and E. granulosus camel strain (camel-dog) in Iran. In Khojir National Park, near Tehran, Iran, a fertile hydatid cyst was recently found in the liver of a dead wild sheep (Ovis orientalis). The number of protoscolices (n=6,000) proved enough for an experimental infection in a dog. The characteristics of large and small hooks of metacestode were statistically determined as the sensu stricto strain but not the camel strain (P=0.5). To determine E. granulosus genotype, 20 adult worms of this type were collected from the infected dog. The second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA (rDNA) and cytochrome c oxidase 1 subunit (COX1) of the mitochondrial DNA were amplified from individual adult worm by PCR. Subsequently, the PCR product was sequenced by Sanger method. The lengths of ITS2 and COX1 sequences were 378 and 857 bp, respectively, for all the sequenced samples. The amplified DNA sequences from both ribosomal and mitochondrial genes were highly similar (99% and 98%, respectively) to that of the ovine strain in the GenBank database. The results of the present study indicate that the morpho-molecular features and characteristics of E. granulosus in the Iranian wild sheep are the same as those of the sheep-dog E. granulosus sensu stricto strain.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권4호
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• pp.234-241
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• 2000

The Saccharomyces cerevisiae PMR1 gene encodes a Ca2+-ATPase localized in the Golgi. We have investigated the effects of PMR1 disruption in S. cerevisiae on the glycosylation and secretion of three heterologous glycoproteins, human ${\alpha}$1-antitrypsin (${\alpha}$1-AT), human antithrombin III (ATHIII), and Aspergillus niger glucose oxidase (GOD). The pmr1 null mutant strain secreted larger amounts of ATHIII and GOD proteins per a unit cell mass than the wild type strain. Despite a lower growth rate of the pmr1 mutant, two-fold higher level of human ATHIII was detected in the culture supernatant from the pmr1 mutant compared to that of the wild-type strain. The pmr1 mutant strain secreted ${\alpha}$1-AT and the GOD proteins mostly as core-glycosylated forms, in contrast to the hyperglycosylated proteins secreted in the wild-type strain. Furthermore, the core-glycosylated forms secreted in the pmr1 mutant migrated slightly faster on SDS-PAGE than those secreted in the mnn9 deletion mutant and the wild type strains. Analysis of the recombinant GOD with anti-${\alpha}$1,3-mannose antibody revealed that GOD secreted in the pmr1 mutant did not have terminal ${\alpha}$1,3-linked mannose unlike those secreted in the mnn9 mutant and the wild type strains. The present results indicate that the pmr1 mutant, with the super-secretion phenotype, is useful as a host system to produce recombinant glycoproteins lacking high-mannose outer chains.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권3호
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• pp.173-178
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• 2003

To improve the erythritol productivity of Penicillium sp. KJ81, mutants were obtained using UV irradiation and NTG treatment Among these mutants, Penicillium sp. KJ-UV29 revealed no morphological changes, yet was superior to the wild strain in the following three points: (1) Penicillium sp. KJ-UV29 produced more erythritol than the wild strain under the same conditions, (2) no foam was produced during cultivation, unlike the wild strain, and (3) the mutant produced a Significantly lower amount of glycerol. Penirillium sp. KJ-UV29 produced as much as 15.1 g/L of erythritol, whereas the wild-type Penirillium sp. KJ81 only produced 11.7 g/L. Penicillium sp. KJ-UV29 only generated 6.1 g/L of glycerol, compared to 19.4 g/L produced by the wild strain. When investigating the optimal culture conditions for erythritol production by the mutant strain Penicillium sp. KJ-UV89, sucrose was identified as the most effective carbon source, and the mutant was even able to produce erythritol in a 70% sucrose-containing medium, although a 30% sucrose medium exhibited the highest productivity. The production of erythritol by Penirillium sp. KJ-UV29 was also significantly increased by the addition of ammonium carbonate, potassium nitrate, and sodium nitrate. Accordingly, under optimal conditions, Penicillium sp. KJ-UV29 produced 45.2 g/L of erythritol in a medium containing 30% sucrose, 0.5% yeast extract, 0.5% (NH$_4$)$_2$C$_2$O$_4$, 0.1% KNO$_3$, 0.1% NaNO$_3$, and 0.01% FeSO$_4$ with 1 vvm aeration and 200 rpm agitation at 37$^{\circ}C$ for 7 days in a 5-L jar fermentor.

• Molecules and Cells
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• 제28권6호
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• pp.575-581
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• 2009

Ornithine decarboxylase (ODC) is a rate-limiting enzyme in the biosynthesis of polyamines, which are essential for cell growth, differentiation, and proliferation. This report presents the characterization of an ODC-encoding cDNA (SlitODC) isolated from a moth species, the tobacco cutworm, Spodoptera litura (Lepidoptera); its expression in a polyamine-deficient strain of yeast, S. cerevisiae; and the recovery in polyamine levels and proliferation rate with the introduction of the insect enzyme. SlitODC encodes 448 amino acid residues, 4 amino acids longer than B. mori ODC that has 71% identity, and has a longer C-terminus, consistent with B. mori ODC, than the reported dipteran enzymes. The null mutant yeast strain in the ODC gene, SPE1, showed remarkably depleted polyamine levels; in putrescine, spermidine, and spermine, the levels were > 7, > 1, and > 4%, respectively, of the levels in the wild-type strain. This consequently caused a significant arrest in cell proliferation of > 4% of the wild-type strain in polyamine-free media. The transformed strain, with the substituted SlitODC for the deleted endogenous ODC, grew and proliferated rapidly at even a higher rate than the wild-type strain. Furthermore, its polyamine content was significantly higher than even that in the wild-type strain as well as the spe1-null mutant, particularly with a very continuously enhanced putrescine level, reflecting no inhibition mechanism operating in the putrescine synthesis step by any corresponding insect ODC antizymes to SlitODC in this yeast system.

• Journal of Microbiology and Biotechnology
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• 제26권11호
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• pp.1908-1917
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• 2016

Wild strain L-6 was subjected to combined mutagenesis, including UV irradiation, atmospheric and room temperature plasma, and ion beam implantation, to increase the yield of 1,3-dihydroxyacetone (DHA). With application of a high-throughput screening method, mutant Gluconobacter oxydans I-2-239 with a DHA productivity of 103.5 g/l in flask-shake fermentation was finally obtained with the starting glycerol concentration of 120 g/l, which was 115.7% higher than the wild strain. The cultivation time also decreased from 54 h to 36 h. Compared with the wild strain, a dramatic increase in enzyme activity was observed for the mutant strain, although the increase in biomass was limited. DNA and amino acid sequence alignment revealed 11 nucleotide substitutions and 10 amino acid substitutions between the sldAB of strains L-6 and I-2-239. Simulation of the 3-D structure and prediction of active site residues and PQQ binding site residues suggested that these mutations were mainly related to PQQ binding, which was speculated to be favorable for the catalyzing capacity of glycerol dehydrogenase. RT-qPCR assay indicated that the transcription levels of sldA and sldB in the mutant strain were respectively 4.8-fold and 5.4-fold higher than that in the wild strain, suggesting another possible reason for the increased DHA productivity of the mutant strain.

• The Plant Pathology Journal
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• 제36권4호
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• pp.355-363
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• 2020

Bacterial traits for virulence of Ralstonia solanacearum causing lethal wilt in plants were extensively studied but are not yet fully understood. Other than the known virulence factors of Ralstonia solanacearum, this study aimed to identify the novel gene(s) contributing to bacterial virulence of R. solanacearum. Among the transposon-inserted mutants that were previously generated, we selected mutant SL341F12 strain produced exopolysaccharide equivalent to wild type strain but showed reduced virulence compared to wild type. In this mutant, a transposon was found to disrupt the murI gene encoding glutamate racemase which converts L-glutamate to D-glutamate. SL341F12 lost its motility, and its virulence in the tomato plant was markedly diminished compared to that of the wild type. The altered phenotypes of SL341F12 were restored by introducing a full-length murI gene. The expression of genes required for flagella assembly was significantly reduced in SL341F12 compared to that of the wild type or complemented strain, indicating that the loss of bacterial motility in the mutant was due to reduced flagella assembly. A dramatic reduction of the mutant population compared to its wild type was apparent in planta (i.e., root) than its wild type but not in soil and rhizosphere. This may contribute to the impaired virulence in the mutant strain. Accordingly, we concluded that murI in R. solanacearum may be involved in controlling flagella assembly and consequently, the mutation affects bacterial motility and virulence.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1392-1398
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• 2006

We investigated a possible coordination between the biosyntheses of two polyketides in the cereal head blight fungus Gibberella zeae, zearalenone (ZEA) and aurofusarin (AUR), which are catalyzed by the polyketide synthases (PKS) PKS4/PKS13 and PKS12, respectively. To determine if the production of one polyketide influences that of the other, we used four different transgenic strains of G zeae; three were deficient for either ZEA or AUR or both, and one was an AUR-overproducing strain. The mycelia of both the wild-type and ${\Delta}PKS4$ strain deficient for ZEA produced AUR normally, whereas the mycelia of both the ${\Delta}PKS12$ and ${\Delta}PKS4::{\Delta}PKS12$ strain showed no AUR accumulation. All the examined deletion strains caused necrotic spots on the surface of com kernels and were found to produce the nonpolyketide mycotoxins trichothecenes to the same amount as the wild-type strain. In contrast, the AUR-deficient ${\Delta}PKS12$ strains produced greater quantities of ZEA and its derivatives than the wild-type progenitor on both a rice substrate and a liquid medium; the AUR-overproducing strain did not produce ZEA on either medium. Furthermore, the expression of both PKS4 and PKS13 was induced earlier in the ${\Delta}PKS12$ strains than in the wild-type strain, and there was no difference in the transcription of PKS12 between the two strains. Therefore, these results indicate that the ZEA biosynthetic pathway is negatively regulated by the accumulation of another polyketide (AUR) in G zeae.

• 미생물학회지
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• 제49권2호
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• pp.195-199
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• 2013

멧돼지 대장에서 MPL-01 균주를 분리하여 균주의 형태학적, 생리 생화적 특성 및 지방산 조성을 분석하여 Bacillus임을 확인하였다. 16S rRNA 유전자 서열 분석 결과 MPL-01 균주는 Bacillus atrophaeus와 거의 일치하므로(99.99%) B. atrophaeus MPL-01로 명명하였다. MPL-01 균주는 고추 탄저병을 일으키는 Colletotrichum acutatum에 대하여 가장 강한 항진균 활성을 나타내었다. 배양액의 ethyl acetate 추출액에서 항진균 활성은 물론이고 계면활성도 확인하였다. 그러므로 B. atrophaeus MPL-01은 농작물 병원성 곰팡이 제어를 위한 bio-control 개발에서 유용하게 사용될 것이다.