得られた酵素調製物を用いたキチナーゼ生産体の産生とミクロミセス細胞壁の破壊

Creation of Chitinase Producer and Disruption of Micromycete Cell Wall with the Obtained Enzyme Preparation.

• O A Sinitsyna
• E A Rubtsova
• I G Sinelnikov
• D O Osipov
• A M Rozhkova
• V Yu Matys
• T V Bubnova
• V A Nemashkalov
• A S Sereda
• L A Tcsherbakova
• A P Sinitsyn

抄録

菌体Penicillium verruculosumをベースに、Myceliophtora thermophilaからキチナーゼを含む細胞外酵素の複合体を生産する組換え株を作製した。生産株の培養液から得られた酵素製剤の活性は、分子量200kDaのキチンと1000kDaのキトサンを用いて、それぞれ0.55、0.53、0.66U/mgのタンパク質を得た。組換えキチナーゼの最適温度は52〜65℃であり、最適pHは4.5〜6.2であり、このクラスの酵素の公表データに対応していた。得られた酵素製剤中の異種キチナーゼの含有量は、培養液中の全タンパク質含有量の47％であった。組換えP. verruculosum XT403株を用いて異種キチナーゼを含む酵素製剤を作製したところ、植物病原性のものを含む微生物の菌糸体を分解することができ、微生物産業廃棄物のバイオコンバージョンにおいて非常に効率的であった。

A recombinant strain producing a complex of extracellular enzymes including chitinase from Myceliophtora thermophila was created based on the fungus Penicillium verruculosum. The activity of the enzyme preparations obtained from the cultural fluid of the producer strain was 0.55, 0.53, and 0.66 U/mg protein with chitin and chitosans with the molecular weight of 200 and 1000 kDa, respectively. The temperature optimum for the recombinant chitinase was 52-65°C; the pH optimum was 4.5-6.2, which corresponded to the published data for this class of the enzymes. The content of heterologous chitinase in the obtained enzyme preparations was 47% of total protein content in the cultural fluid. Enzyme preparations produced by the recombinant P. verruculosum XT403 strain and containing heterologous chitinase were able to degrade the mycelium of micromycetes, including phytopathogenic ones, and were very efficient in the bioconversion of microbiological industry waste.