• Journal of Microbiology and Biotechnology
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• 제16권2호
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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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• 제34권2호
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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.

• 한국식품영양학회지
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• 제15권4호
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• pp.364-369
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• 2002

방선균은 토양 속에 다양하게 존재하는 미생물의 일종으로 그람 양성 진정세균으로 이차대사산물을 생산하는 시기와 포자 착생이 시작되는 세포분화의 시기가 밀접한 관련이 있다. S. griseus는 streptomycin을 비롯한 다양한 종류의 endopeptidase 및 exopeptidase들을 생산한다. 방선균에서의 protease 생산은 많은 경우에 이차대사산물이 형성되거나 형태분화가 유도되는 시기에 동시에 시작된다는 점에서 Pretense가 이차대사물질 생산 및 세포분화에 일정한 기능을 수행할 것이 라는 점을 시사하고 있다. 본 연구에서는 S. griseus IFO 13350에서 클로닝한 SGPD protease가 각 strain에서 형태학적으로나 생리적으로 어떠한 gene dosage 효과를 미치는지 조사하는 것이었다. sprD 유전자가 S.lividans를 숙주로 사용한 시스템에서 대량발현이 성공적으로 되는 것을 확인한 후, 본 유전자를 클로닝한 S. griseus IFO13350 균주와 이의 A-factor 결손주인 S. griseus HH1에 형질전환하였다. S. griseus HH1과 S. griseus IFO13350에서는 protease activity가 벡터만 도입된 대조군과 sprD 유전자가 들어간 형질전환체에서 큰 차이를 보이지 않았다. 또한 S. griseus IFO 13350 및 HH1 모두에서 생리학적·형태학적 분화의 차이를 발견하지 못하였다. Chymotrypsin계열의 pretense를 암호화하는 유전자만이 S. griseus에서 발현이 repression된다는 사실을 본 연구 결과를 통하여 알게 되었다. 이를 바탕으로 sprD유전자와 동일계열의 chymotrypsin 계열의 유전자들이 공통적으로 S. griseus에서 repression 되는 일반적인 기전이 있을 것으로 판단, chymotrypsin계열 유전자들의 promoter부분의 염기 상동성을 조사하였다 번역개시부위 바로 상부 유전자부터 상동성을 조사한 결과 적어도 상당부분의 염기배열이 잘 보존된 지역이 존재함을 알게 되었다. 향후 이들 발현기구의 조절기구를 연구함으로서 protease의 기능을 밝히는데 좋은 단서를 제공할 것으로 판단된다.

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

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

• Journal of Microbiology and Biotechnology
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• 제15권5호
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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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• 제39권4호
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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.

• 한국항공우주학회지
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• 제39권5호
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• pp.442-450
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• 2011

회전 블레이드의 모션을 측정하기 위해 스테레오 패턴 인식 (stereo pattern recognition, SPR) 기법을 적용한 측정 시스템을 구성하고 실험을 수행하였다. 시스템 요구사항을 만족시키는 스테레오 카메라를 확보하여 SPR 기반의 측정 시스템을 준비하였다. 시스템의 측정불확도를 계산하기 위한 일련의 실험을 통해, 본 SPR 시스템이 2m~4.2m의 측정 거리에서 0.2mm 이내의 표준불확도를 가짐을 확인하였다. 이를 이용하여 계산한 블레이드 모션의 합성표준불확도는, 리드-래그, 플래핑, 비틀림 운동에 대해 각각 0.296mm, 0.209mm, $0.238^{\circ}$ 이었다. 본 SPR 시스템을 한국항공우주연구원의 소형로터 시험장치에 설치하고 회전속도와 콜렉티브 피치각을 각각 360rpm, 589rpm, 그리고 $0^{\circ}$, $4^{\circ}$, $6^{\circ}$로 바꾸어가며, 지상 제자리비행 조건에서 블레이드의 모션과 탄성 변형을 성공적으로 측정하였다. Higher Harmonic Control Aeroacoustic Rotor Test -II 에서 사용한 측정 시스템과의 비교를 통하여 본 시스템의 장점을 분석하였다.

• Journal of Welding and Joining
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• 제33권2호
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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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• 제11권6호
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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.