Use of bacilli for overproduction and secretionof endo-β-1,4-glucanase and β-glucosidase encoded by cloned genes = 크론된 유전자에 의한 endo-β-1,4-glucanse와 β-glucosidase 의 Bacillus 숙주를 통한 과잉 생산 및 분비

A substantial overproduction and secretion of endo-$\beta$-1,4-glucanase via a shuttle vector, pCK98, carrying a Bacillus subtilis gene coding for this enzyme was allowed in $\underline{Bacillus}$ $\underline{subtilis}$ and $\underline{B.}$ $\underline{megaterium}$. Activities of the exocellular enzymes produced by $\underline{B.}$ $\underline{subtilis}$ RM125 (pCK98) and $\underline{B.}$ $\underline{megaterium}$ (pCK98) were found to be higher than that produced by the original gene donor $\underline{B.}$ $\underline{subtilis}$ strain by 52- and 43-fold, respectively. Most of the enzyme was produced during the exponential growth period and the production was not repressed by glucose or cellobiose. The plasmid was stable in $\underline{B.}$ $\underline{megaterium}$ but not in $\underline{B.}$ $\underline{subtilis}$. The production of $\beta$-glucosidase via new shuttle vectors having a $\underline{Alcaligenes}$ $\underline{faecalis}$ gene encoding this enzyme was less effectively allowed in $\underline{Bacillus}$ and $\underline{E.}$ $\underline{coli}$ cells even though the gene was put on amplifiable plasmids. Contrary to the endo-$\beta$-1, 4-glucanase gene, the $\beta$-glucosidase gene was comparatively unstable in Bacillus and E. coli cells. On the analysis of plasmids isolated from resultant transformed cells, intactness, deletion, rearrangement or modification of $\beta$-glucosidase gene portion in shuttle plasmids were all found. The concomitant production of glucose from carboxymethyl cullulose (CMC) with $\beta$-glucosidase and endo- $\beta$-1,4-glucansase complex produced by $\underline{Bacillus}$ or $\underline{E.}$ $\underline{coli}$ transformants was observed depending on the comparatively long incubation period. Additionally, a novel procedure using polysaccharides and dyes for the rapid detection and comparison of various polysaccharase activities was established. The starch, CMC, lichenan, laminarin, or pustulan as substrates, together Congo Red...
Pack, Moo-Youngresearcher박무영researcher
Issue Date
61005/325007 / 000825217

학위논문(박사) - 한국과학기술원 : 생물공학과, 1987.2, [ xii, 125 p. ]

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