Culture Characteristics on the Activity of ${\gamma}-Glutamyltranspeptidase{\;}({\gamma}-GTP)$ by Bacillus subtilis Fusant

배양조건이 Bacillus subtilis 융합주의 ${\gamma}-Glutamyltranspeptidase{\;}({\gamma}-GTP)$ 활성에 미치는 영향

  • Published : 2001.06.01

Abstract

A fusant FG-21 was selected on the basis of higher ${\gamma}-GTP$ activity following fusion process between SM-2 and SM-10 of Bacillus subtilis mutants. ${\gamma}-GTP$ activity of the mutant FG-21 was increased up to 612 U/mL when grown for 36 hr at $37^{\circ}C$ in culture media containing 1% glycerol 1% glycerol, 1% peptone, 0.1% citric acid, 5 mM $K_2HPO_4$, 1 mM $FeCl_3$, 1 mM $MgCl_2$, 1 mM $NH_4Cl$, pH 7.0. In fusnat FG-21, the ratio of protein to total sugar contents for biopolymer A was 38 to 59. for biopolymer B from parental strains it was 19 to 78. Fructose contents determined by HPLC were $573.7\;\mu\textrm{g}/mg\;and\;764.4\;\mu\textrm{g}/mg$ for biopolymer A and B, respectively. And glutamic acid content were $163.7\;\mu\textrm{g}/mg\;and\;94.6\;\mu\textrm{g}/mg$ for biopolymer A and B, respectively. In fusant FG-21, the ratio of fructose to glutamic acid contents for biopolymer A was 78 to 22. For biopolymer B from parental strains it was 89 to 11.

Bacillus subtilis 돌연변이주 중 ${\gamma}-GTP$활성이 높은 SM-2와 SM-10을 융합시켜 획득한 융합주 중 ${\gamma}-GTP$활성이 높은 융합주 FG-21의 배양적 특성을 조사하였다. 융합주 FG-21에 의한 ${\gamma}-GTP$의 생산은 1% glycerol, 1% peptone, 0.1% citric acid, 5 mM $K_2HPO_4$, 1 mM $FeCl_3$, 1 mM $MgCl_2$, 1 mM $NH_4Cl$, pH 7.0의 배지에 의해 $37^{\circ}C$에서 36시간 배양했을 때 621 U/mL으로 최대한 활성을 보였다. 융합주 FG-21이 생산한 biopolymer A가 단백질함량이 38.4%이고 B. subtilis K-1이 생산하는 biopolymer B의 단백질함량은 19.3%로 융합주 FG-21이 생산하는 Biopolymer A보다 함량이 낮았다. 융합주 FG-21의 biopolymer가 모균주의 biopolymer B보다 단백질함량이 높은 것은 상대적으로 높은 ${\gamma}-GTP$활성이 높아서 생성된 PGA가 levan보다 함량이 많기 때문인 것으로 사료된다. 총당함량은 biopolymer A가 58.5%, biopolymer B가 76.5%로 나타났다. 융합주 FG-21과 모균주가 생산하는 biopolymer의 단백질함량과 총당함량비는 융합주 FG-21이 생성한 biopolymer는 38 : 59. 모균주가 생산한 biopolymer B는 19 : 78의 비율이었고 HPLC 분석에 의한 biopolymer A와 B의 fructose함량은 각각 537.7 mg/g biopolymer, 764.4 mg/g biopolymer으로 나타났다. Biopolymer A와 B의 glutamic acid함량은 각각 163.7 mg/g, 94.3 mg/g이었다. Biopolymer A와 B의 fructose와 glutamic acid의 함량비는 각각 78 : 22, 89 : 11의 비로 함유되어 있었다. 따라서 융합주에 의해 생산된 biopolymer는 모균주보다 구성상태가 변화된 biopolymer를 생산한 것으로 판단되어 fusion과 같은 균주개량을 통하여 물질의 구성적, 물질적 변화된 biopolymer를 생산할 수 있을 것이라 사료된다.

Keywords

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