• Journal of Microbiology and Biotechnology
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• v.14 no.6
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• pp.1350-1355
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• 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.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2011.06a
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• pp.1321-1322
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• 2011

• Transactions of the Korean Society of Mechanical Engineers B
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• v.30 no.1 s.244
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• pp.8-15
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• 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
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• v.11 no.6
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• pp.1077-1086
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• 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.

• Korean Journal of Microbiology
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• v.41 no.1
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• pp.87-92
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• 2005

Pronase, a protease produced for commercial purpose by Streptomyces griseus, was composed of serine protease, alkaline protease, aminopeptidase and carboxypeptidase complex, and it has been widely used as anti-inflammatory drugs for human therapy. In this study, we developed a new integration vector, pHJ101 derived from pSET152, containing strong promoter, ermE, to overexpress a certain protease gene. Specific PCR primers for cloning of sprA (a gene for S. griseus protease A) and sprB (a gene for S. griseus protease B) genes were designed from the basis of nucleotide sequence in databases and amplified by PCR. Plasmid pHJ201 and pHJ202 were constructed by inserting of amplified each gene in a vector pHJ101. S. griseus HA and S. griseus HB were respectively obtained by conjugal process of a parent strain, S. griseus IFO 13350 with the recombinant Escherichia coli harboring plasmid pHJ201 or pHJ202. When protease activity was measured in flask cultivation, produced protease levels of S. griseus HA and S. griseus HB increased about 5.3 times and 5 times, respectively, more than that of parent strain. And, the constructed integrating plasmid pHJ101 was applicable for overexpression of a certain gene in Streptomyces sp.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1125-1129
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• 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
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• v.24 no.11
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• pp.1595-1600
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• 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.

• Transactions of the Korean Society of Mechanical Engineers A
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• v.38 no.7
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• pp.735-742
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• 2014

A self-piercing rivet (SPR) is a mechanical component for joining dissimilar material sheets such as those of aluminum alloy and steel. Unlike conventional rivets, the SPR directly pierces sheets without the need for drilling them beforehand. However, the regular SPR can undergo buckling when it pierces a high-strength steel sheet, warranting the design of a helical SPR. In this study, the joining and forging processes using the helical SPR were simulated using the commercial FEM code, DEFORM-3D. High-tensile-strength steel sheets of different strengths were joined with aluminum alloy sheets using the designed helical SPR. The simulation results were found to agree with the experimental results, validating the optimal design of a helical SPR that can pierce high-strength steel sheets.

• BMB Reports
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• v.31 no.1
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• pp.101-105
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• 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
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• v.12 no.2
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• pp.167-172
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• 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.