• Biotechnology and Bioprocess Engineering:BBE
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• 제11권2호
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• pp.127-133
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• 2006

Shewanella oneidensis MR-1 has the ability to inhale certain metals and chemical compounds and exhale these materials in an altered state; as a result, this microorganism has been widely applied in bioremediation protocols. However, the relevant characteristics of cell growth and biosynthesis of PuFAs have yet to be thoroughly investigated. Therefore, in this study, we have attempted to characterize the growth and fatty acid profiles of S. oneidensis MR-1 under a variety of temperature conditions. The fastest growth of S. oneidensis MR-1 was observed at $30^{\circ}C$, with a specific growth rate and doubling time of $0.6885h^{-1}\;and\;1.007 h$. The maximum cell mass of this microorganism was elicited at a temperature of $4^{\circ}C$. The eicosapentaenoic acid (EPA) synthesis of S. oneidensis MR-1 was evaluated under these different culture temperatures. S. oneidensis MR-1 was found not to synthesize EPA at temperatures in excess of $30^{\circ}C$, but was shown to synthesize EPA at temperatures below $30^{\circ}C$. The EPA content was found to increase with decreases in temperature. We then evaluated the EPA biosynthetic pathway, using a phylogenetic tree predicted on 16s rRNA sequences, and the homology of ORFs between S. oneidensis MR-1 and Shewanella putrefaciens SCRC-2738, which is known to harbor a polyketide synthase (PKS)-like module. The phylogenetic tree revealed that MR-1 was very closely related to both Moritella sp., which is known to synthesize DHA via a PKS-like pathway, and S. putrefaciens, which has been reported to synthesize EPA via an identical pathway. The homology between the PKS-like module of S. putrefaciens SCRC-2738 and the entire genome of S. oneidensis MR-1 was also analyzed, in order to mine the genes associated with the PKS-like pathway in S. oneidensis MR-1. A putative PKS-like module for EPA biosynthesis was verified by this analysis, and was also corroborated by the experimental finding that S. oneidensis MR-1 was able to synthesize EPA without the expression of $dihomo-{\gamma}-linoleic$ acid (DGLA) and arachidonic acid (AA) formed during EPA synthesis via the FAS pathway.

• Asian-Australasian Journal of Animal Sciences
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• 제30권12호
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• pp.1784-1795
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• 2017

Objective: The study examined the effect of intravenous administration of bacterial endotoxin-lipopolysaccharide (LPS) -on the nocturnal secretion of melatonin and on the expression of enzymes of the melatonin biosynthetic pathway in the pineal gland of ewes, taking into account two different photoperiodic conditions: short-night (SN; n = 12) and long-night (LN; n = 12). Methods: In both experiments, animals (n = 12) were randomly divided into two groups: control (n = 6) and LPS-treated (n = 6) one. Two hours after sunset, animals received an injection of LPS or saline. Blood samples were collected starting one hour after sunset and continuing for 3 hours after the treatment. The ewes were euthanized 3 hours after LPS/saline treatment. The concentration of hormones in plasma was assayed by radioimmunoassay. In the pineal gland, the content of serotonin and its metabolite was determined by HPLC; whereas the expression of examined genes and protein was assayed using real-time polymerase chain reaction and Western Blot, respectively. Results: Endotoxin administration lowered (p<0.05) levels of circulating melatonin in animals from LN photoperiod only during the first hour after treatment, while in ewes from SN photoperiod only in the third hour after the injection. Inflammation more substantially suppressed biosynthesis of melatonin in ewes from SN photoperiod, which were also characterised by lower (p<0.05) cortisol concentrations after LPS treatment compared with animals from LN photoperiod. In the pineal gland of ewes subjected to SN photoperiod, LPS reduced (p<0.05) serotonin content and the expression of melatonin biosynthetic pathway enzymes, such as tryptophan hydroxylase and arylalkylamine-N-acetyltransferase. Pineal activity may be disturbed by circulating LPS and proinflammatory cytokines because the expression of mRNAs encoding their corresponding receptors was determined in this gland. Conclusion: The present study showed that peripheral inflammation reduces the secretion of melatonin, but this effect may be influenced by the photoperiod.

• Journal of Plant Biotechnology
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• 제29권2호
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• pp.139-144
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• 2002

Sterols play two major roles in plants: a bulk component in biological membranes and precursors of plant steroid hormones. Physiological effects of plant steroids, brassinosteroids (BRs), include cell elongation, cell division, stress tolerance, and senescence acceleration. Arabidopsis mutants that carry genetic defects in BR biosynthesis or its signaling display characteristic phenotypes, such as short robust inflorescences, dark-green round leaves, and sterility. Currently there are more than 100 dwarf mutants representing 7 genetic loci in Arabidopsis. Mutants of 6 loci, dwf1/dim1/cbb1, cpd/dwf3, dwf4, dwf5, det2/dwf6, dwf7 are rescued by exogenous application of BRs, whereas bri1/dwf2 shares phenotypes with the above 6 loci but are resistant to BRs. These suggest that the 6 loci are defective in BR biosynthesis, and the one locus is in BR signaling. Biochemical analyses, such as intermediate feeding tests, examining the levels of endogenous BR, and molecular cloning of the genes revealed that dwf7, dwf5, and dwf1 are defective in the three consecutive steps of sterol biosynthesis, from episterol to campesterol via 5-dehydroepisterol. Similarly, det2/dwf6, dwf4, and cpd /dwf3 were shown to be blocked in D$^4$reduction, 22a-hydroxylation, and 23 a-hydroxylation, respectively. A signaling mutant bril/dwf2 carries mutations in a Leucine-rich repeat receptor kinase. Interestingly, the bri1 mutant was shown to accumulate significant amount of BRs, suggesting that signaling and biosynthesis are dynamically coupled in Arabidopsis. Thus it is likely that transgenic plants over-expressing the rate-limiting step enzyme DWF4 as well as blocking its use by BRIl could dramatically increase the biosynthetic yield of BRs. When applied industrially, BRs will boost new sector of plant biotechnology because of its potential use as a precursor of human steroid hormones, a novel lead compound for cholesterol-lowering effects, and a various application in plant protection.

• 미생물학회지
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• 제41권4호
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• pp.300-305
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• 2005

Corynebacterium glutamicum에서 promoter-probe vector인 pSK1Cat을 이용해 분리된 프로모터를 함유하는 단편들 중 가장 높은 활성을 나타낸 $P_{19}$ 단편에 대한 심도 있는 분석을 수행하였다. Subcloning을 실시하여 프로모터 활성을 지닌 DNA 영역을 180 bp로 압축할 수 있었고 $(P_{180})$, 이를 C. glutamicum의 균주개량 측면에서 그 활용성을 분석하였다. C. glutamicum에서 메치오닌 생합성에 관여하는metX유전자의 메치오닌에 의한 repression을 해제시키기 위하여 metX유전자의 promoter를 $P_{180}$ promoter로 교체하였고 $(P_{180}-metX)$, $P_{180}-metX$를 C. glutamicum에 도입하여 발현되는 homoserine acetyltransferase 활성을 다양한 성장조건에서 측정하였다. MB 영양배지에서 배양하는 경 우 $P_{180}-metX$를 함유는 균주는 wild type보다 약 24배 높은 homoserine acetyltransferase 활성을 나타내었다. Tac 프로모터에 연계하는 경우 $(P_{tac}-metX)$, 약 13배의 활성 증가만이 관찰되었다. 최소배지에서 배양한 후 분석한 결과, $P_{180}-metX$에서의 발현양상은 배지에 첨가된 methionine에 의해 영향받지 앓음을 확인하였는데, 이는 $P_{180}$ 단편이 생합성 유전자의 derepression에 의한 아미노산 생산균의 개량에 효율적으로 이용될 수 있음을 의미한다. $P_{180}-metA$를 라이신 생산균에 도입하는 경우 최대 약 0.8g/l의 메치오닌이 생산됨을 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권6호
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• pp.812-819
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• 2018

Objective: An experiment was conducted to identify and characterize the circular RNA expression and metabolic characteristics in the liver of Jinhua pigs and Landrace pigs. Methods: Three Jinhua pigs and three Landrace pigs respectively at 70-day were slaughtered to collect the liver tissue samples. Immediately after slaughter, blood samples were taken to detect serum biochemical indicators. Total RNA extracted from liver tissue samples were used to prepare the library and then sequence on HiSeq 2500. Bioinformatic methods were employed to analyze sequence data to identify the circRNAs and predict the potential roles of differentially expressed circRNAs between the two breeds. Results: Significant differences in physiological and biochemical traits were observed between growing Jinhua and Landrace pigs. We identified 84,864 circRNA candidates in two breeds and 366 circRNAs were detected as significantly differentially expressed. Their host genes are involved in lipid biosynthetic and metabolic processes according to the gene ontology analysis and associated with metabolic pathways. Conclusion: Our research represents the first description of circRNA profiles in the porcine liver from two divergent phenotype pigs. The predicted miRNA-circRNA interaction provides important basis for miRNA-circRNA relationships in the porcine liver. These data expand the repertories of porcine circRNA and are conducive to understanding the possible molecular mechanisms involved in miRNA and circRNA. Our study provides basic data for further research of the biological functions of circRNAs in the porcine liver.

• BMB Reports
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• 제42권11호
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• pp.752-757
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• 2009

Copper is essential but toxic in excess for aerobic organisms. Yeast transcription factor ACE1 functions as a sensor for copper and an inducer for the transcription of CUP1. In addition, ACE1 can activate the transcription of superoxide dismutase gene (sod1) in response to copper. In this study, we introduced the yeast ACE1 into Arabidopsis and analyzed its function in plant. Under high copper stress, the transgenic plants over-expressing ACE1 showed higher survival rate than the wild-type. We also found that over-expression of ACE1 in Arabidopsis increased the activities of SOD and POD, which were beneficial to the cell in copper buffering. Excess copper would suppress the expression of chlorophyll biosynthetic genes in Arabidopsis, RT-PCR analysis revealed that over-expression of ACE1 decrease the suppression. Together, our results indicate that ACE1 may play an important role in response to copper stress in Arabidopsis.

• Molecules and Cells
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• 제41권4호
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• pp.311-319
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• 2018

The gaseous hormone ethylene influences many aspects of plant growth, development, and responses to a variety of stresses. The biosynthesis of ethylene is tightly regulated by various internal and external stimuli, and the primary target of the regulation is the enzyme 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS), which catalyzes the rate-limiting step of ethylene biosynthesis. We have previously demonstrated that the regulation of ethylene biosynthesis is a common feature of most of the phytohormones in etiolated Arabidopsis seedlings via the modulation of the protein stability of ACS. Here, we show that various phytohormones also regulate ethylene biosynthesis from etiolated rice seedlings in a similar manner to those in Arabidopsis. Cytokinin, brassinosteroids, and gibberellic acid increase ethylene biosynthesis without changing the transcript levels of neither OsACS nor ACC oxidases (OsACO), a family of enzymes catalyzing the final step of the ethylene biosynthetic pathway. Likewise, salicylic acid and abscisic acid do not alter the gene expression of OsACS, but both hormones downregulate the transcript levels of a subset of ACO genes, resulting in a decrease in ethylene biosynthesis. In addition, we show that the treatment of the phytohormones results in distinct etiolated seedling phenotypes, some of which resemble ethylene-responsive phenotypes, while others display ethylene-independent morphologies, indicating a complicated hormone crosstalk in rice. Together, our study brings a new insight into crosstalk between ethylene biosynthesis and other phytohormones, and provides evidence that rice ethylene biosynthesis could be regulated by the post-transcriptional regulation of ACS proteins.

• 한국미생물·생명공학회지
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• 제13권1호
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• pp.19-25
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• 1985

Saccharomyces cerevisiae HIS 5 유전자는 histidinol phosphate aminotransferase (EC: 2, 6, 1,9)를 code하는 아미노산 합성유전자이다. 이 유전자는 plasmid pSH 530에 cloning되어 E. coli와 Saccharomyces cerevisiae 숙주에서 promoter로서 전사하였다. HIS 5 유전자의 총염기 수는 736개이였고 5' 상류영역에는 긴 reading frame, directed repeat, 전사개시점, 그리고 Pribnow box염기배열이 있었다. 특히 HIS 5 유전자의 ATG 주변 염기배열은 -A-A-A-T-T-A-C-A-C-T-A-T-G-G-T-T-T-T-T-G-A-T-였으며 C block은 없었다.

• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1801-1809
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• 2015

A phytoene synthase gene, crtB, was isolated from Kocuria gwangalliensis. The crtB with 1,092 bp full-length has a coding sequence of 948 bp and encodes a 316-amino-acids protein. The deduced amino acid sequence showed a 70.9% identity with a putative phytoene synthase from K. rhizophila. An expression plasmid, pCcrtB, containing the crtB gene was constructed, and E. coli cells containing this plasmid produced the recombinant protein of approximately 34kDa , corresponding to the molecular mass of phytoene synthase. Biosynthesis of lycopene was confirmed when the plasmid pCcrtB was co-transformed into E. coli containing pRScrtEI carrying the crtE and crtI genes encoding lycopene biosynthetic pathway enzymes. The results obtained from this study will provide a base of knowledge about the phytoene synthase of K. gwangalliensis and can be applied to the production of carotenoids in a non-carotenoidproducing host.

• Journal of Applied Biological Chemistry
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• 제54권4호
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• pp.244-251
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• 2011

Galactinol and rafinose accumulation in plants is associated with stressful environmental conditions such as cold, heat, or dehydration by the action of galactinols synthase (GolS) in the raffinose family of oligosaccharides biosynthetic pathway from UDP-galactose. Moreover, several reports mentioned that GolS transcription is up regulated by various environmental stresses like cold, heat, dehydration. Therefore, to determine whether MoGolS1 was induced with the abiotic stress we analyzed the expression pattern of the gene under various abiotic stresses like heat, cold, abscisic acid, sucrose and salt concentration in the lemon balm plants grown in standard MS medium. The MoGolS1 gene was 981-bp in length encoding 326 amino acids in its sequence and shared 77 and 76% sequence similarity with Arabidopsis thaliana galactinol synthase4 (AtGolS4) and AtGolS1 genes respectively. The MoGolS1 gene was strongly expressed by the abiotic stress induced by sucrose, ABA or heat shock. It was also expressed in responses to cold, Identification and Functional Characterization of the GALACTINOL SYNTHASgene induction with various stresses may be possible for itscrucial function in abiotic stress tolerance in plants, providing a good engineering target for genetic engineering.