• Journal of Microbiology and Biotechnology
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• v.16 no.2
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• pp.312-317
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• 2006

The chromosomal sprD gene encoding Streptomyces griseus protease D (SGPD), a chymotrypsin-like protease, was disrupted in Streptomyces griseus by insertion of the neomysin-resistance gene. The production of chymotrypsin activity of sprD disruptant was not completely abolished, but delayed by 24 h, compared with that of wild-type strain. The aerial mycelial formation of sprD disruptant was retarded, and specifically the formation of spores was not observed in the central region of colonies. However, normal morphological development into spores was observed in the marginal region of colonies. In addition, the production of yellow pigment that might be dependent on A-factor was also decreased in the sprD disruptant, compared with that of the wild-type strain. Introduction of the sprD gene, which was placed on a high copy-numbered plasmid into S. griseus ${\Delta}sprD$, partially restored the ability of morphological development, and a significant level of sporulation was observed. When the overexpression vector for sprD, pWHM3-D, was introduced in S. griseus, there was no significant change in the chymotrypsin activity or colonial morphology, in contrast to Streptomyces lividans, indicating the presence of a tight regulation system for the overexpression of the sprD gene in S. griseus.

• Journal of Welding and Joining
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• v.34 no.2
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• pp.54-58
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• 2016

SPR(Self-Piercing rivet) is mechanical element of joining sheet metal components without the need for pre-punched or pre-drilled holes. Newly designed SPR is developed for high joining strength and shearing strength than semi-tubular rivet. In this study, divided shank of self-piercing rivet were designed for joining DP440 and SILAFONT. Newly designed SPR was simulated by using FEM code DEFORM-3D. In simulations of SPR process, various shape of self-piercing rivet were considered for semi-tubular and newly designed SPR. In other to examine the joinability, joining load and lap-shear load of newly designed SPR were compared with semi-tubular by simulated results and experimental ones.

• The Korean Journal of Food And Nutrition
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• v.15 no.4
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• pp.364-369
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• 2002

Streptomyces shows a eukaryotic characteristic that vegetative cell can grow into mycelial form and has morphological and physiological differentiation at a certain period during its life cycle. Streptomyces has been used for the production of many biologically active compounds, such as antibiotics and pronase. Production of second metabolites and differentiation of the vegetative cell share the certain period of its lift cycle. Therefore, second metabolites may affect the differentiation of the vegetative cell. One of the microbial hormone, called A-factor, regulates the production of second metabolites, sporulation and differentiation of the cells. Streptomyces griseus produces streptomycin as well as many different kinds of proteinase. As mentioned, period of proteinases production overlaps with the period of differentiation of the vegetative cells. Protease may play a important role for the differentiation of the cells. In this paper, function of the SGPD gene cloned from S. griseus IFO 13350 tested whether it affects for the differentiation of A-factor mutated S. griseus HH1 and S. griseus IFO13350. pWHM3 and pWHM3-sprD plasmid was transformed into S. griseus HH1 and S. griseus IFO13350. Chymotrypsin activity of the cultured medium of the transformants with pWHM3-sprD plasmid didn't show any change with that of the transformants with plasmid only. The transformants with pWHM3-sprD plasmid didn't show the increase of the production of actinorhodin as well as morphological change in S. griseus IFO 13350 and HH1, as well. The promoter sequences of the SGPA and SGPB gene which encode chymotrypsin-like protease, were compared with that of SGPD gene. Regulatory mechanism of gene expression of proteinase genes will be studied for the development of high production system for protease as well as the function of the proteases.

• 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.

• 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

• 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.

• Journal of Microbiology
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• v.39 no.4
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• pp.305-313
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• 2001

The spr D gene encoding Strptomyces griseus protease D(SGPD); a chymotrypsin-like proteae, was cloned from Strptomyces griseus IFO13350 and sequence. Most of the amino-acid sequence deduced from the nucleotide sequence is idential to that Strptomyces griseus IMRU3499 except that one amino acid has been deleted and Trp 369 has been substituted into Cys369 in the SGPD from S. griseus IFO13350 without affecting the protease activity. The spr D gene was overexpressed in Streptomyce liv-idans TK24 as a heterologous host. Various media with different compositions were also used to max-imize the productivity of SGPD inthe heterologous host. The SGPD productivity was best when the transformant S. lividans TK24 was cultivated in R2YE medium. The relative chymotrypsin activity of the culture broth measured with an artificial chromogenic substrate, N-scuccinyl-ala-ala-pro-phe-p-nitroanilide, was 16 units/ml. A high level of SGPD was also produced in YEME and SAAM medial but it was relatively lower that in R2YE medium and negligible amounts of SGPD were produced in GYE, GAE and Benedict media. The growth of S. lividans reacted the maximum level of cell mass at days 3 and 4 of the culture, but SGPD production started in the stationary phase of cell growth and kept increase in till the 10$^{th}$ day of culture in R2YE and YEME medium, but in GYE media the productivity reached maximum level at 8days of cultivation. The introduction of the spr D gene into S. lividans TK24 triggered biosyntheis of the pigmented antibiotic , actinorhodin, which implies some protease may paly a very improtant role in secondary-metabolite formation in sStreptomyces.

• Journal of the Korean Society for Aeronautical & Space Sciences
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• v.39 no.5
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• pp.442-450
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• 2011

A measurement system using stereo pattern recognition (SPR) method was configured to measure the rotor blade deformations and motions. An SPR-based measurement system was prepared using six stereo cameras. Through a series of experiments to evaluate the system measurement uncertainty, it was verified that the SPR system had less than 0.2mm standard uncertainty. The combined standard uncertainties for the lead-lag, flapping, and pitching motions were estimated as 0.296mm, 0.209mm, and $0.238^{\circ}$, respectively. The SPR system was installed at a general small-scaled rotor test system at Korea Aerospace Research Institute. The blade motions and elastic deformation were successfully measured under the conditions with rotating speeds of 360rpm or 589rpm, and collective pitch angles of $0^{\circ}$, $4^{\circ}$, or $6^{\circ}$. The advantages of the SPR system was analyzed in comparison with the measurement system used in Higher Harmonic Control Aeroacoustic Rotor Test -II.

• Journal of Welding and Joining
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• v.33 no.2
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• pp.78-84
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• 2015

Self-piercing rivet(SPR) is mechanical joining methods and which can be joining dissimilar materials. Unlike conventional riveting, SPR also needs no pre-drilled holes. During plastically deformation, SPR pierces upper sheet and joins it to under sheet. SPR has been mainly applied to the joining the automobile body and some materials, such as glass fiber reinforced polymer and aluminum alloy, which represent the sheet-formed materials for lightweight automobile. Glass fiber reinforced plastic(GFRP) has been considered as a partial application of the automobile body which is lighter than steels and stronger than aluminium alloys. It is needed SPR to join Al alloy sheets and GFRP ones. In this paper, in order to design the rivet and anvil, which are suitable for GFRP, the joinability was examined through simulations of SPR joining between GFRP and Al alloy sheets. For this study, AutoCAD was used for the modeling and the simulated using commercial FEM code DEFORM-2D. The simulated results for SPR process joining between GFRP and Al alloys were confirmed by the same conditions as experimental trials.

• 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.