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RISS 인기검색어
- 검색키워드
- 저자 : Sinskey, Anthony J. (검색결과 12 건)
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Molecular Characterization of AceB, a Gene Encoding Malate Synthase in Corynebacterium glutamicum
LEE, HEUNG-SHICK,SINSKEY, ANTHONY J. 한국미생물 · 생명공학회 1994 Journal of microbiology and biotechnology Vol.4 No.4
The aceB gene, encoding for malate synthase, one of the key enzymes of glyoxylate bypass, was isolated from a pMT1-based Corynebacterium glutamicum gene library via complementation of an Escherichia coli aceB mutant on an acetate minimal medium. The aceB gene was closely linked to aceA, separated by 598 base pairs, and transcribed in divergent direction. The aceB expressed a protein product of Mr 83,000 in Corynebacterium glutamicum which was unusually large compared with those of other malate synthases. A DNA-sequence analysis of the cloned DNA identified an open-reading frame of 2,217 base pairs which encodes a protein with the molecular weight of 82,311 comprising 739 amino acids. The putative protein product showed only limited amino acid-sequence homology to its counterparts in other organisms. The N-terminal region of the protein, which shows no apparent homology with the known sequences of other malate synthases, appeared to be responsible for the protein's unusually large size. A potential calciumbinding domain of EF-hand structure found among eukaryotes was detected in the N-terminal region of the deduced protein.
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세포외 다당류 생산을 위한 Zoogloea ramigera 115 균주의 개량
임무현,D. Davidson Easson Jr,Anthony J. Sinskey 대구대학교 산업기술연구소 1989 産業技術硏究 Vol.8 No.-
세포의 다당류의 생산능이 우수한 균주를 Zoogloea ramigera 115로부터 변이 처리하여 선발하였다. Z. ramigera 115균이 Capsule을 형성하여 floc을 만드는데 반하여 이 균은 slime층을 형성하여 용액중에 잘풀리는 다당류를 생산하는 특징을 보여주었으며, Z. ramigera115 SL로 명명하였다. 따라서 이균은 배양적 성질이 모균인 Z. ramigera에 비하여 세포외 다당의 분리가 용이하게 개량되었고 모균에 비하여 약 20%의 생산성의 향상을 나타내었다. 이들 두 균주가 생산하는 다당의 구성분석에서 glucose와 galactose의 구성비가 약 2:1로 모두 동일하게 나타났으며 Z. ramigera 115 SL의 경우 Pyruvate와 D-acetyl group의 함량이 모균주보다 약간 높은 수치를 보였다.
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Cloning and Sequencing of the Gene Involved in Morphological Chang of Zoogloea ramigera 115SLR
Lee, Sam Pin,Kim, Tae Rahk,Anthony John Sinskey 한국미생물 · 생명공학회 2000 Journal of microbiology and biotechnology Vol.10 No.2
Plasmid pLEX3 isolated from the recombinant cosmid library of Zoogloea ramigera 115 was found to be responsible for the restoration of the rugose colony phenotype. To confirm the essential region responsible for the complementation, subclones were constructed from plasmid pLEX3 and transformed into mutant strain Z ramigera 115SLR. The recombinant plasmids pLEX10 and pLEX11 were shown to complement the slime-forming property of Z. ramigera 115SLR. In a compositional analysis of the exopolysaccharides from Z ramigera 115, Z ramigera 115SLR, and Z ramigera 115SLR harboring plasmid pLEX11, the exopolysaccharides showed a similar composition with glucose, galactose, and side chain groups. The complete nucleotide sequence of the 3.25kb genomic DNA insert in plasmid pLEX11 was determined and its analysis identified two open reading frames which could encode two proteins. The gene products derived from the two open reading frames were confirmed by an in vivo transcription using a T7-RNA polymerase. The ORF1 produced a 30kDa protein, whereas the ORF2 was found responsible for the complementation of the morphological mutation and produced a 14kDa protein. An in vivo gene expression of plasmid pTEX10 showed another open reading frame encoding a 50kDa protein. The gene products from ORF1 and ORF2 are regarded as novel proteins which do not show any homology with other proteins.
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양영헌,전종민,이다혜,김정호,서형민,Cho Kyun Rha,Anthony J. Sinskey,Christopher J. Brigham 한국생물공학회 2015 Biotechnology and Bioprocess Engineering Vol.20 No.2
Conventional solvent-based methods are still the most practical approaches for recovery of polyhydroxyalkanoate (PHA) polymer from cellular biomass, even though potential alternatives exist, including chemical, mechanical, and enzymatic methods. It is still necessary, however, to avoid dangerous and environmentally unfriendly solvents (e.g., chloroform and dichloromethane) in the polymer recovery process. In the work presented here, we applied various solvent systems to recover PHA from Ralstonia eutropha and recombinant Escherichia coli cells. It was demonstrated that methyl ethyl ketone (MEK) is a promising solvent for PHA recovery from bacterial cells, particularly for the copolymer poly(hydroxybutyrate-cohydroxyvalerate) [P(HB-co-HV)], exhibiting > 90% polymer recovery. Even though MEK did not solubilize PHAs to the same extent as chloroform, it can recover a comparable amount of polymer because of its processing advantages, such as the low viscosity of the MEK/PHA solution, and the lower density of MEK as compared to cellular components. MEK was found to be the best alternative, non-halogenated solvent among examined candidates for recovery of P(HB-co-HV) from cells. The MEK treatment of PHAcontaining cells further allowed us to eliminate several costly and lengthy steps in the extraction process, such as cell lysis, centrifugation, and filtration.
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