• Journal of Plant Biology
• /
• 제36권1호
• /
• pp.91-96
• /
• 1993

Phycoerythrins from the ceramiaceous red algae Campylaephora crassa (Okamura) Nakamura and related species, C. hypnaeoides J. Agardh and Ceramium kondoi Yendo, were investigated for absorption spectra, protein bands by gel electrophoresis and antigenic reactivity to anti-phycoerythrin using Ouchterlony double diffusion and immunoblot. Similarities in absorption spectra, showing peaks at ca. 566 nm>534 nm>495 nm, were found between C. crassa and Cm. kondoi, while C. hypnaeoides differed slightly. There were no differences in fluorescence emission spectra and protein bands between C. crassa and related species tested. Since Ouchterlony double diffusion, however, showed that phycoerythrins from C. crassa and Cm. kondoi were similar in antigenic reactions, and differed from that of C. hypnaeoides, the taxonomic position of C. crassa should be reinvestigated using other experimental approaches.

• ALGAE
• /
• 제20권4호
• /
• pp.325-331
• /
• 2005

Populations of Campylaephora borealis (Nakamura) Seo, Cho et Boo and C. crassa (Okamura) Nakamura show a year-around occurrence of all life-history stages. Such a concurrency of life-history stages produces problems in recognizing species in the field. Here, we invesitgated the morphological variation and life-history stages of both species using a statistical character analysis. Life-history stage was correlated with the seawater temperature in C. borealis, whereas it was dependant on biomass in C. crassa. Thalli had dichotomous branches with adaxial branchlets. The statistics showed that the seasonal change in morphology of C. borealis was significantly different from that of C. crassa in seven qualitative characters and five quantitative characters (p < 0.001), although six quantitative features including tetrasporangial size were similar in both species. The morphological difference between the two species may be due to the annual variation of branchlet number and the variance of branch subangle.

• 미생물학회지
• /
• 제26권2호
• /
• pp.73-81
• /
• 1988

$\alpha$-Amylase (EC.3.2.1.1) of Neurospora crassa (ATCC9279) was cloned in E. coli HB101 using shotgun method, and the enzymes isolated from both N. crassa and E. coli were compared. Chromosomal DNA isolated from the spores of N. crassa was partially digested with PstI restriction endonuclease and rejoined to pBR322 which had been digested with the same enzyme. The resulting recombinant DNA were introduced into E. coli HB101 which had competancy by treating with $CaCl_{2}$. As the result, about 8000 colonies which showed tetracycline resistance were selected and two of the colonies which had 13.5Kb recombinant plasmid exhibit starch degrading activity on starch-containing plate when treated with D-cycloserine. $\alpha$-Amylases from both N.crassa and E. coli were isolated by using ammonium sulfate precipitation, DEAE-cellulose ion exchange column chromatography and Bio-Gel P150 gel foltration column. As the result, about 81.3 fold and 5.6 fold purifications in specific activities were obtained respectively, and specific activities of the gel filtrates were 6.1u/mg and 85u/mg respectively. The properties of both enzymes were compared and they showed quite the similar patterns in optimal temperature, optimal pH and had same molecular weight about 100,000 daltons on gel filtration method. Optimal temperatures for both enzymes were $70^{\circ}C$ and optimal pH were about 6 and 10.

• 생명과학회지
• /
• 제13권6호
• /
• pp.933-942
• /
• 2003

Coenzyme Q은 긴 isoprenoid 사슬을 갖는 quinone의 유도체이다. Coenzyme Q는 진핵생명체의 미토콘드리아의 내막과 원핵생명체의 세포막에 위치하는 전자전달계에 존재하는 지용성 물질이며, 또한 항산화제로의 기능도 갖는다. Coenzyme Q는 Saccharomyces cerevisiae의 호기적 성장에 필수적이며, coq 돌연변이체는 발효가 불가능한 탄소 원에서의 성장이 불가능하다. S. cerevisiae의 $coq^7$p 효소들과 유사성을 나타내는 단백질을 암호화하는 Neurcspora crassa cDNA를 효모의 발현 벡터에 삽입하였다. N. crassa COQ7의 예상 서열은 S. cerevisiae의 효소와 58% homology를 보였다. N. crassa $coq^{-7}$ 유전자의 S. cerevisiae $coq^7$ 형질전환체는 야생형 균주와 유사한 성장률을 보였다. 형질전환 균주들은 발효가 불가능한 탄소원인 글리세롤을 유일한 탄소원으로 배양하였을 경우에도 정상적인 성장을 나타냈다. 또한 불포화지방산인 linolenic acid를 성장 배지에 첨가하여도 야생형 균주와 유사한 생존율이 관찰되었다.

• Journal of Microbiology and Biotechnology
• /
• 제27권9호
• /
• pp.1664-1669
• /
• 2017

Gamma-aminobutyric acid is a precursor of nylon-4, which is a promising heat-resistant biopolymer. GABA can be produced from the decarboxylation of glutamate by glutamate decarboxylase. In this study, a synthetic scaffold complex strategy was employed involving the Neurospora crassa glutamate decarboxylase (GadB) and Escherichia coli GABA antiporter (GadC) to improve GABA production. To construct the complex, the SH3 domain was attached to the N. crassa GadB, and the SH3 ligand was attached to the N-terminus, middle, and C-terminus of E. coli GadC. In the C-terminus model, 5.8 g/l of GABA concentration was obtained from 10 g/l glutamate. When a competing pathway engineered strain was used, the final GABA concentration was further increased to 5.94 g/l, which corresponds to 97.5% of GABA yield. With the introduction of the scaffold complex, the GABA productivity increased by 2.9 folds during the initial culture period.

• 미생물학회지
• /
• 제13권2호
• /
• pp.45-50
• /
• 1975

Uptake of $^{14}C$-sorbose and $^{14}C$-3-O-methylglucose by ungerminated conidia of Neurospora crassa was measured by means of the millipore filter technique. Initial rates of jptake of both sorbose and 3-O-methylglucose show a marked dependence optimal pH for uptake of both sugars is close to 4.75. When ungerminated conidia are "starved" with buffer for a prolonged period of time prior to assaying their transport capacity and mycelia, no de-repression of the glucose-repressible sugar transport system is effectuated in contrast to the findings for germinated conidia.d conidia.

• 미생물학회지
• /
• 제41권4호
• /
• pp.246-254
• /
• 2005

Neurospora crassa의 게놈 분석을 통하며 tetratricopeptide repeat (TPR) 부위를 보유할 것으로 추정되는 19중의 단배질을 찾아냈다. 이중에서 한 단백질은 Saccharomyces cerevisiae에서 다양한 유전자들의 공통 전사 억제인자로 알려진 Ssn6 단백질에 $60\%$ 이상의 유사도를 보였다. N. crassa의 RIP (repeat-induced point mutation) 과정을 통하여 생산된 돌연변이 균주들은 도두 느리게 성장하였는데, 4가지로 구분되는 돌연변이 표현형을 나타냈다. 첫 번째 돌연변이 표현형은 균사가 ropy돌연변이와 유사한 균사 모양으로 성장하고 밀도가 높아 보였고, 대분생포자는 yellow와 csp 표현형을 보였다. 두 번째 돌연변이형은 늦은 성장을 보였지만 대분생포자를 생산하였다. 세 번째는 매우 느린 균사 성장을 보였고 acon 표현형을 나타냈다. 마지막 돌연변이형은 거의 공기 중으로 균사를 뻗지 못하였으며 acon 표현형을 보였는데, rco-1 RIP 돌연변이체와 유사하였다. 돌연변이체들은 모두 male로서는 수정능을 보였지만 female로서는 교배가 불가능했으며 자낭각을 생산하지 못하였다. 이 결과들은 이 유전자가 N. crassa의 성장은 물론 무성생식 및 유성생식의 여러 과정에 관여함을 나타낸다. 염색체와 cDNA의 서열을 분식한 결과에 따르면, 유전자가 6개의 인트론을 보유하고 있고, 총917개의 아미노산으로 구성된 102kDa의 단백질을 암호화할 것으로 예상되었다. 이 유전자를 rcm-1 (regulation of conidiation and morphology)으로 명명하였다.

• Journal of Microbiology and Biotechnology
• /
• 제16권9호
• /
• pp.1468-1471
• /
• 2006

The last step in L-carnitine biosynthesis in eukaryotic organisms is mediated by ${\gamma}$-butyrobetaine hydroxylase (EC1.14.11.1), a dioxygenase that converts ${\gamma}$-butyrobetaine to L-carnitine. This enzyme was previously identified from rat liver and humans, and the peptide sequence of human ${\gamma}$-butyrobetaine hydroxylase was used to search the Neurospora crassa genome database, which led to an identification of an open reading frame (ORF) consisting of 1,407 bp encoding a polypeptide of 468 amino acids. When this protein was expressed in Saccharomyces cerevisiae, the crude cell-free extract exhibited ${\gamma}$-butyrobetaine hydroxylase activity.

• 한국어병학회지
• /
• 제2권1호
• /
• pp.1-18
• /
• 1989

지금까지 조사보고(調査報告)된 바 없는 한국산(韓國産) 양식(養殖) 뱀장어의 부레에 기생(寄生)한 선충에 의(依)한 피해여부(被害與否)와 예방대책(豫防對策)을 수립할 목적(目的)으로 기생(寄生)한 선충의 종(種)과 기생율(寄生率), 병해성(病害性) 그리고 생활사(生活史)를 조사(調査)하였다. 1. 성숙(成熟)한 충란의 크기는 $80{\sim}92(86.7){\times}62}\sim}71(67.4){\mu}m$였고, 자충의 크기는 $210{\sim}240(225){\times}18{\sim}23(20.6){\mu}m$였다. 2. 본충의 중간숙주(中間宿主)는 Thermocyclops hyalinus이며, 이것의 복강(腹腔)에 침입(侵入)한 유충은 4일후(日後)에 감염력(感染力)을 가졌고 이때의 유충 크기는 $360{\sim}420(390){\times}28{\sim}35(31)\;{\mu}m$였다. 3. 감염력(感染力)을 갖는 Thermocyclops hyalinus를 뱀장어가 섭취(攝取)한 뒤 50일(日) 경과(經過)되어 뱀장어 부레내에서 성충으로 발육(發育)되었다. 4. 검출(檢出)된 성충의 크기는 female$7.3{\sim}31.0(16.5){\times}0.5{\sim}2.2(1.2)mm$, male$4.9{\sim}13.3(8.3){\times}0.3{\sim}0.9(0.4)mm$로서 내외부(內外部) 형웅(形熊)를 비교(比較)한 바 Anguillicola crassa로 동정(同定)하였다. 5. 뱀장어에서의 월별기생율(月別奇生率)은 6, 9, 12월(月)에 높았고, 1~3월(月)에 비교적(比較的) 낮았다. 6. 감염(感染)되지 않은 정상어(正常魚)와 감염어(感染魚)의 비만도(肥滿度)를 월별(月別), 체장별(體長別)로 그 유의성(有意性)을 검정(檢定)한 결과(結果) 유의성(有意性)을 인정(認定)할 수 없었으며 본충 감염(感染)으로 인(因)한 뱀장어 성장(成長)에는 영향(影響)이 없는 것 같았다. 7. 본충의 기생(寄生)에 의한 뱀장어 부레조직(組織)의 이상(異狀)은 보이지 않았다. 8. 수온(水溫) $25.0{\sim}26.7^{\circ}C$ 때 masoten을 처리(處理)한 농도별(濃度別) Thermocyclops hyalonus의 사망시간(死亡時間)은 0.5ppm 14시간(時間), 0.4ppm 20시간(時間), 0.3ppm 22시간(時間), 0.2ppm 30시간(時間), 0.1ppm 42시간(時間)이었다.

• Archives of Pharmacal Research
• /
• 제8권3호
• /
• pp.149-157
• /
• 1985

The HCL hydrolyzate of the non-histone protein fractionated from the rat liver nuclei which have been incubated inthe presence of S-adenosyl-L-[methyl-$^{14}C$ ]-methionine shows at least four unidentified radioactive peaks on a basic amino acid analysis chromatogram. One of these unknown compounds (designated as compound 3) is also formed by the rat liver homogenated with the exogenous addition of an appropriate protein substrate. Since boiled rat liver homogenate or fresh homogenate in the absence of an exogenous protein substrate failed to form compound 3, its formation can be considered to be enzyme-catalyzed. The enzyme which yields compound 3 shows a preference of protein substrate in the order of reductively methylated hemoglobin > native > histone type II-A. The rat enzyme is nuclear in location associated with chromatin, and exhibits the highest activity in the liver among various rat organs. A compound 3-forming enzyme is also present in Neurospora crassa, since endogenous formation of the compound 3 can be demonstrated with the crude extract of this mold. The chemical identity of compound 3 is not yet known. However, it resisted to the following treatments; 6 N HCL and 0.1 N Na NaOH hydrolysis at $110^{\circ}C$, OR L-amino acid oxidase.