• Journal of Microbiology and Biotechnology
• /
• 제14권6호
• /
• pp.1350-1355
• /
• 2004

The sprA and sprB genes, encoding the chymotrypsin-like proteases Streptomyces griseus protease A (SGPA) and Streptomyces griseus protease B (SGPB), and the sprT gene that encodes Streptomyces griseus trypsin (SGT) were cloned from S. griseus and were overexpressed in various strains of S. griseus. When the sprT gene was introduced into S. griseus, trypsin activity increased 2-fold in the A-factor deficient mutant strain, S. griseus HH1, and increased 4-fold in the wild strain, S. grise us IFO 13350. However, there was no detectable increase of chymotrypsin activity in the transformants of S. griseus with either sprA or sprB, in contrast to the results obtained from S. lividans as a heterologous host. To solve the negative gene dosage effects in S. griseus, either the sprA or the sprB genes with their own ribosome binding sites were linked to the downstream of the entire sprT gene, and the coexpression efficiency was examined in S. lividans and S. griseus. The transformants of S. lividans with either pWHM3-TA (sprT+sprA) or pWHM3­TB (sprT+sprB) showed 3-fold increase of trypsin activity over that of the control, however, only the transformant of pWHM3-TB demonstrated 7-fold increase in chymotrypsin activity, indicating that the pWHM3-TB has a successful construction for the overexpression of chymotrypsin in Streptomyces. When the coexpression vectors were introduced into S. griseus IFO 13350, the trypsin level sharply increased by more than 4-fold, however, the chymotrypsin level did not increase. These results strongly suggest that the overexpression of the sprA and sprB genes is tightly regulated in S. griseus.

• 한국정밀공학회:학술대회논문집
• /
• 한국정밀공학회 2011년도 춘계학술대회 논문집
• /
• pp.1321-1322
• /
• 2011

• 대한기계학회논문집B
• /
• 제30권1호
• /
• pp.8-15
• /
• 2006

A Sparse Point Representation(SPR) based on interpolation wavelets is presented. The SPR is implemented for the purpose of CFD data compression. Unlike conventional wavelet transformation, the SPR relieves computing workload in the similar fashion of lifting scheme that includes splitting and prediction procedures in sequence. However, SPR skips update procedure that is major part of lifting scheme. Data compression can be achieved by proper thresholding method. The advantage of the SPR method is that, by keeping even point physical values, low frequency filtering procedure is omitted and its related unphysical thresholing mechanism can be avoided in reconstruction process. Extra singular feature detection algorithm is implemented for preserving singular features such as shock and vortices. Several numerical tests show the adequacy of SPR for the CFD data. It is also shown that it can be easily extended to nonlinear adaptive wavelets for enhanced feature capturing.

• Journal of Microbiology and Biotechnology
• /
• 제11권6호
• /
• pp.1077-1086
• /
• 2001

The sprA and sprB gene encoding chymotrypsin-like proteases Streptomyces griseus protease A (SGPA) and Streptomyces griseus protease B (SGPB) and the sprT gene that encodes Streptomyces griseus trypsin (SGT) were cloned from Streptomyces griseus ATCC10137 and overexpressed in Streptomyces lividans TK24 as a heterologous host. The chymotrypsin activity of tole culture broth measured with the artificial chromogenic substrate , N-succinyl-ala-ala-pro-phe-p-nitroanilide, was 10, 14 and 14 units/mg in the transformants haboring the sprA, sprB and sprD genes, respectively. The growth of S. lividans reached the maximum cell mass after 4 days of culture, yet SGPA and SGPD production started in the stationary phase of cell growth and kept increasing for up to 10 days of culture in an R2YE medium. The trypsin activity of the culture broth measured with the artificial chromogenic substrate , N-${\alpha}$-benzoyl-DL- arginine-p-nitroanilide , was 16 units/mg and SGT production started in the stationary phase of cell growth and kept increasing for up to 10 days of culture in an R2YE medium. The introduction of the sprA gene into S, lividans TK24 triggered the biosynthesis of pigmented antibiotics, actinorhodin and undecylprodigiosin, and induced significant morphological changes in the colonies in Benedict, R2YE, and R1R2 media. In addition, the introduction of the sprT gene also induced morphological changes in the colony shape without affecting the antibiotic production, thereby implying that certain proteases would appear to play very important and specific roles in secondary-metabolites formation and morphological differentiation in Streptomyces.

• 미생물학회지
• /
• 제41권1호
• /
• pp.87-92
• /
• 2005

방선균 Streptomyces griseus에서 상업적 목적으로 생산되는 protease인 protease는 serine protease, alkaline protease, aminopeptidase및 carboxypeptidase로 구성되어 있는 복합체로서 의 약용 소염제로 널리 사용되어지고 있다. 본 연구에서는 기존에 개발되어 있는 방선균용 integration vector인 pSET152로부터 목적산물의 대량발현을 위해 방선균용 promoter ermE가 cloning된 새로운 integration vector인 pHJ101을 개발하였고, pretense의 생산량 증대에 사용하였다. 새로 개발된 integration vector에 S. griseus protease A를 코드하고 있는 유전자, sprA와 S. griseus pretense B유전자, sprB를 각각 cloning하여 plasmid pHJ201과 pHJ202를 구축하였다. 이들 plasmid들을 S. griseus IFO 13350에 형질전환하여 발현용plasmid가 chromosome에 integration된 재조합 균주 S. gliseus HA와 S. griseus HB를 얻었다. 이들 재조합균주로부터 전체 protease의 생산량을 확인한 결과, 모균주보다 각각 S. griseus HA는 약 5.3 배, S. griseus HB는 약 5 배 정도 생산량이 증대되었다. 이들 결과로부터 특정유전자의 고발현용 integration vector의 제작이 확인되었으며, 전체 protease의 생산량 증대의 가능성이 시사되었다.

• Journal of Microbiology and Biotechnology
• /
• 제15권5호
• /
• pp.1125-1129
• /
• 2005

Cloning of a 6.6-kb BamHI digested chromosomal DNA from S. griseus IFO13350 revealed the presence of an additional gene encoding a novel trypsin-like enzyme, named SprU. The SprU protein shows a high homology ($79\%$ identity, $88\%$ similarity) with the SGT protease, which has been reported as a bacterial trypsin in the same strain. The amino acid sequence deduced from the nucleotide sequence of the sprU gene suggests that SprU is produced as a precursor consisting of an amino-terminal presequence (29 amino acid residues), prosequence (4 residues), and mature trypsin consisting of 222 amino acids with a molecular weight of 22.94 kDa and a calculated pI of 4.13. The serine, histidine, and aspartic acid residues composing the catalytic triad of typical serine proteases are also well conserved. When the trypsin activity of the SprU was spectrophotometrically measured by the enzymatic hydrolysis of the artificial chromogenic substrate, N-${alpha}$-benzoyl-DL-arginine-p-nitroanilide, the S. lividans transformant with pWHM3-U gave 3 times higher activity than that of control. When the same recombinant plasmid was introduced into S. griseus, however, the gene dosage effect was not so significant, as in the cases of other genes encoding serine proteases, such as sprA, sprB, and sprD. Although two trypsins, SprU and SGT, have a high degree of homology, the pI values, the gene dosage effect in S. griseus, and the gene arrangement adjacent to the two genes are very different, suggesting that the biochemical and biological function of the SprU might be quite different from that of the SGT.

• Asian-Australasian Journal of Animal Sciences
• /
• 제24권11호
• /
• pp.1595-1600
• /
• 2011

Sugarcane press residue (SPR), a byproduct from the sugar industry was evaluated for it's nutrient and energetic quality in broilers and layers. The composition of SPR included (% DM): CP-11.76 (methionine-2.21, cystine-1.05, lysine-4.85, threonine-5.48% of CP), EE-7.87 (palmitic acid-30.3, stearic acid-4.1, oleic aicd-17.2, linoleic acid-38.0, linolenic acid-5.4% of EE), CF-10.08, TA-21.08 (Ca-3.87, P-1.10, Mg-0.95%, Fe-3500, Mn-284, Zn-113, Cu-61.5, Co-5.0 ppm and AIA-4.93%) and NFE-48.35% indicating that SPR is a valuable source of both organic and inorganic nutrients for poultry. The metabolic trials revealed the average ME of SPR as 749, 842 and 1,270 kcal/kg, respectively in broilers and 844, 936 and 1,031 kcal/kg in layers, at 10, 20 and 30% inclusion levels, respectively. Further, the fortification of SPR incorporated diets with biotechnological products viz., lipid utilizing agents (lipase and lecithin) or NSP degrading enzymes and their combination did not improve the ME content of such diets.

• 대한기계학회논문집A
• /
• 제38권7호
• /
• pp.735-742
• /
• 2014

Self-piercing rivet(SPR)은 이종재료 접합을 위해 사용되는 결합용 기계요소로써, 대표적으로는 알루미늄 합금과 강판 등 용융점이 서로 다른 재료의 접합에 사용된다. SPR 접합은 일반 리벳접합과 달리 스스로 홀을 가공하며 삽입되기 때문에 사전의 홀 가공이 필요 없다.(1) 상부판재를 천공하고 하부판재와 함께 소성 변형되어 결합된다. 자동차의 차체 경량화를 위해서는 알루미늄 합금과 같은 경량소재가 사용되며, 부분적으로 스틸과 알루미늄 합금의 이종재료 접합이 요구된다. 그러나 알루미늄 합금과 강판은 용융점이 다르므로 기존의 차체 결합방법으로 이용되고 있는 저항 용접이 불가능하다. 이에 따라, 기계적 결합방법의 하나인 SPR 접합이 요구된다.(2) 따라서 본 연구에서는 강소성 유한요소해석 프로그램을 이용하여 리벳과 판재의 접합 성형성을 검토하고, 고장력 강판을 접합할 수 있는 새로운 형상의 SPR을 설계하였다. 또한 해석결과와 실험의 비교를 통하여 해석의 신뢰성을 검증하였다.

• BMB Reports
• /
• 제31권1호
• /
• pp.101-105
• /
• 1998

Immunoassay of botulinum toxin (BTX) B type was investigated using two typed of biosensors: light addressable potentiometric sensor (LAPS) and surface plasmon resonance (SPR) sensor. Urease-tagged and immuno-filtration capture method have been used for LAPS. Tag-free and direct binding real-time detection method have been used for SPR sensor. The detection limit of sandwich assay format with LAPS was 10 ng/ml, which was the lowest among methods tested. SPR has the advantage of being more convenient because tag-free direct binding assay can be used and reaction time was reduced, regardless of low sensitivity. This result shows that sandwich assay format with LAPS can be used as an alternative method of BTX mouse bioassay which is known as the most sensitive method for the detection of BTX.

• Journal of Electrochemical Science and Technology
• /
• 제12권2호
• /
• pp.167-172
• /
• 2021

he progressively improved sensing sensitivity (∆λ_SPR/∆n, nm/RIU) to detect the refractive index is observed on the SPR platform of an Au-covered epoxy gratings in an increase in potential cycling in a typical three-electrode cell. Here, a DVD-R optical disc was used as a structure template to prepare an Au-covered epoxy gratings, and the newly formed reverse track pitch structure on the epoxy substrate was used as a working electrode directly in aqueous sulfuric acid solution. It is expected that Au reconstruction by potential cycling in sulfuric acid electrolyte increases the packing density of Au atoms in the grain boundary and improves the propagation of electromagnetic waves.