• 미생물학회지
• /
• 제26권1호
• /
• pp.6-12
• /
• 1988

Cellobiase ($\beta$-glucosidase) is an enzyme of the cellulase system in cellulolytic microor-ganisms. The chromosomal DNA fragment which include cellobiase gene of Cellulomonas biazotea was cloned in Eschericia coli via plasmid pBR 322 vector. Restriction enzyme Sal I was used to obtain adequate size of fragments from C. biazotea. chromosomal DNA. The transformant of E. coli HB101 with recombinant plasmid pBG101 showed cellobiase activity, which is not ordinary in E. coli HB101. The enzyme activity of the transformant was as of 20% lower than that of C. biazotea.

• 생명과학회지
• /
• 제23권4호
• /
• pp.542-553
• /
• 2013

본 연구의 목적은 통계학적 방법을 사용하여 해양미생물 Cellulophaga lytica LBH-14가 생산하는 cellobiase의 생산조건을 확립하는 것이었다. 이 균주의 생육에 최적인 미강, ammonium chloride 및 배지의 초기 pH는 100.0 g/l, 5.00 g/l 및 7.0이었으나, 이 균주가 생산하는 cellobiase의 생산에 최적인 조건은 각각 91.1 g/l, 9.02 g/l 및 6.6이었다. 이 균주의 생육에 최적인 $K_2HPO_4$, NaCl, $MgSO_4{\cdot}7H_2O$ 및 $(NH_4)_2SO_4$ 등과 같은 배지의 염농도는 각각 6.25, 0.62, 0.28 및 0.73 g/l이었으나, cellobiase 생산에 최적인 염들의 농도는 각각 4.46, 0.36, 0.27 및 0.73 g/l이었다. 또한, 균체의 생육 및 cellobiase의 생산에 최적인 온도는 각각 35 및 $25^{\circ}C$이었다. 플라스크 규모에서 최적화한 조건으로 파이롯트 규모의 생물배양기에서 cellobiase를 생산한 결과, 이 균주가 생산하는 cellobiase의 생산성은 92.3 U/ml이었으며, 이는 최적화하기 전에 비하여 5.4배 향상된 것 이었다. 본 연구를 통하여 쌀 도정공정의 부산물인 미강 및 ammonium chloride를 cellobiase를 생산하는 기질로 개발하였으며 해양 미생물을 사용하여 cellobiase의 생산기간을 7일에서 3일로 단축시켰다. 또한, 본 연구를 통하여 C. lytica LBH-14가 생산하는 cellobiase의 최적 생산조건은 이 균주가 생산하는 CMCase의 최적 생산조건과 다르다는 사실을 확인하였다.

• 한국미생물생명공학회:학술대회논문집
• /
• 한국미생물생명공학회 1986년도 추계학술대회
• /
• pp.524.1-524
• /
• 1986

CMCase, a member of cellulose decomposing enzymes, hydrolyze cellulose up to cellobiose. Cellobiase splits cellobiose to glucose units. Therefore, a linkage of the twogenes coding for CMCase and cellobiase on the same plasmid is needed to produce a cellulase complex which can produce glucose from cellulose. A genetic operon in which the two structural genes are under the control of a single promoter would be ideal for this purpose. The present report is on the linking of the two cellulase genes in one plasmid as a preliminary step of the operon construction.

• Journal of Microbiology and Biotechnology
• /
• 제14권6호
• /
• pp.1196-1199
• /
• 2004

A bacterium, isolated from rabbit's waste and identified as Cellulomonas sp., had cellulase and thermostable $\alpha$-amylase activity when grown on wheat bran. Maximum activity of thermostable $\alpha$-amylase was obtained by adding $3\%$ soluble starch. However, soybean oil (1 ml $1^{-1}$) could increase the production of $\alpha$-amylase and cellulase in 'wheat bran. The $\alpha$-amylase was characterized by making a . demonstration of optimum activity at $90^{\circ}C$ and pH 6- 9, with soluble starch as a substrate. The effect of ions on the activity and the stability of this enzyme were investigated. This strain secreted carboxymethyl cellulase (CMCase), cellobiase ($\beta$­glucosidase), and filter paperase (Fpase) during growth on wheat bran. Carboxymethy1cellulase, cellobiase, and Fpase activities had pH optima of 6, 5.5, and 6, respectively. CMCase and cellobiase activities both had an optimum temperature of $50^{\circ}C$, whereas Fpase had an optimum temperature of $45^{\circ}C$.

• Journal of Microbiology and Biotechnology
• /
• 제21권4호
• /
• pp.412-420
• /
• 2011

Secretion of cellobiase occurred in a brefeldin A (BFA) uninhibited manner in the filamentous fungus Termitomyces clypeatus. Fluorescence confocal microscopy revealed that application of the drug at a concentration of 50 ${\mu}g$/ml caused arrest of Spitzenkorper assembly at the hyphal tip. This resulted in greater than 30% inhibition of total protein secretion in the culture medium. However, the cellobiase titer increased by 17%, and an additional 13% was localized in the vacuolar fraction en route secretion. The secretory vacuoles formed in the presence of the drug were also found to be bigger (68 nm) than those in the control cultures (40 nm). The enzyme secreted in the presence and absence of BFA revealed a single activity band in both cases in native PAGE and had similar molecular masses (approx. 120 kDa) in SDS-PAGE. The BFA enzyme retained 72% of native glycosylation. It also exhibited a higher stability and retained 98% activity at $50^{\circ}C$, 93.3% activity at pH 9, 63.64% activity in the presence of 1M guanidium hydrochloride, and 50% activity at a glucose concentration of 10 mg/ml in comparison to 68% activity, 75% activity, 36% activity, and 19% activity for the control enzyme, respectively. The observations collectively aimed at the operation of an alternative secretory pathway, distinct from the target of brefeldin A, which bypassed the Golgi apparatus, but still was able to deliver the cargo to the vacuoles for secretion. This can be utilized in selectively enhancing the yield and stability of glycosidases for a successful industrial recipe.

• 미생물학회지
• /
• 제22권3호
• /
• pp.167-173
• /
• 1984

The cellabiase (${\beta}$-glucosidase) gene in a Cellulomonas sp. CS1-1 was cloned into E. coli HB101 using the vector plasmid pBR322, and the expression of the gene in E. coli studied. The chromosomal DNA of the cellulomonas was digested by seveal restriction enzymes, each of which has only one cleaving site in plasmid pBR322. The recombinant plasmid, pSB2, created with Sal I frament, was expressed for the cellobiase gene in E. coli. The recombiant plasmid was estimated to contain 6.4 Kb foreign DNA at the Sal I site of plasmid pBR322 and the inserted DNA was mapped by single and double digestion with several enzymes. E. coli HB101(pSB2) has slowly grown in a mineral liquid medium containing cellobiose as a sole carbon source. The cellobiase activity in the transformed E. coli was 132 units per liter, which is equivalent to one twenty fifth of that in doner strain Cellulomonas sp. CS1-1. The transforned cell with plasmid containing cellulase gene grow well in the LB mediuns. The synthesis of cellobiase in the strain, E. coli HB101 (pSB2), was inhibited by glucose and at high concentration of cellobiose, and induced by cellobiose at low concentration.

• 미생물과산업
• /
• 제11권1호
• /
• pp.21-33
• /
• 1985

이글은 다음과 같이 구성되어져 있다. preface 1) introduction 2) general information 3) folin protein determination 4) cellobiase assay 5) filter paper assay for saccharifying cellulase 6) carboxymethyl cellulase assay for endo-.betha.-1,4-glucanase 7) additional assay procedure for endoglucanase 8) evalutaiton of cellulase under process conditions 9) general remartks, references.

• 한국미생물·생명공학회지
• /
• 제25권2호
• /
• pp.115-120
• /
• 1997

Penicillium sp.-L4, a causative fungus of rot in citrus fruits, was isolated and its mode of hydrolytic enzyme production was investigated. Carboxymethylcellulase (CMCase), polygalacturonase(PGase), extra- & intra-cellular $\beta$-glucosidase and cellobiase were produced drastically by addition of substrates in minimal media. Production of the hydrolytic enzymes were induced efficiently by cellobiose and cellooligosaccharides which were the products of cellulose hydrolysis, but repressed by addition of mono-saccharide such as glucose, raffinose, galacturonic acid. The relative activity of p-nitrophenyl-$\beta$-D-glucopyranoside(PNPG) hydrolysis was higher than that of cellobiose hydrolysis in extracellular enzymes, and reverse is true in intracellular enzymes. Intact enzyme production of P. sp.-L4 on lemon peel lesion was sequential. $\beta$-Glucosidase and CMCase were produced first and followed by PGase. The enzyme productivities and pH in lesions were coincident with optimal pH of each enzyme activities.

• Journal of Microbiology and Biotechnology
• /
• 제25권7호
• /
• pp.1101-1107
• /
• 2015

The role of CRE1 in a thermophilic fungus, Myceliophthora thermophila ATCC42464, was studied using RNA interference. In the cre1-silenced strain C88, the filter paper hydrolyzing activity and β-1,4-endoglucanase activity were 3.76-, and 1.31-fold higher, respectively, than those in the parental strain when the strains were cultured in inducing medium for 6 days. The activities of β-1,4-exoglucanase and cellobiase were 2.64-, and 5.59-fold higher, respectively, than those in the parental strain when the strains were cultured for 5 days. Quantitative reverse-transcription polymerase chain reaction showed that the gene expression of egl3, cbh1, and cbh2 was significantly increased in transformant C88 compared with the wild-type strain. Therefore, our findings suggest the feasibility of improving cellulase production by modifying the regulator expression, and an attractive approach to increasing the total cellulase productivity in thermophilic fungi.

• 한국미생물·생명공학회지
• /
• 제9권2호
• /
• pp.65-69
• /
• 1981

농산폐자원을 이용하기 위한 방법으로써 Trichoderma sp. KI 7-2로 만든 koji와 내열성 효모를 사용하여 동시당화-발효와 당화액 발효를 비교하였다. 볏짚의 당화액에서는 15 mg/$m\ell$의 cellobiose가 존재하였으나 동일한 효소원을 사용한 동시당화-발효액에서는 존재하지 않았다. 효소당화법에서 는 cellulase enzyme system의 반응산물인 glucose가 cellobiose의 활성을 저해하므로 cellobiose 가 축적되었으나 동시당화-발효법에서 는 glucose가 ethanol로 발효되어 cellobiose의 축적이 없었다. Cutting mill 한 볏짚은 동시 당화-발효 과정에서는 ball mill 한 것과 같은 정도로 효과적으로 발효되었다. 이 결과로부터 cellylolytic enzyme system의 반응산물에 의한 저해 mechanism을 논의하였다. 또한 볏짚 당화시 약 10 mg/$m\ell$ 생산되는 xylose는 동시당화-발효에 아무런 영향을 미치지 않음을 확인하였다.