Purification and Characterization of Carboxymethyl Cellulase IV from Penicillium verruculosum

Penicillium verruculosum 으로부터 Carboxymethyl Cellulase IV 의 정제(精製) 및 특성(特性)

An endo-type cellulase, carboxymethyl cellulase(CMCase) IV, was purified from culture filtrate of cellulolytic fungus Penicillium verruculosum. The CMCase IV was acidic glycoprotein having carbohydrate of 13% as glucose and pI value of 4.0. The CMCase IV was 52 KDa of molecular weight in SDS-polyacrylamide gel electrophoresis and have optimum temperature and pH of $50^{\circ}C$ and 5.0 for enzyme activity. The CMCase IV liberated glucose and cellobiose as major products of the enzyme against carboxymethyl cellulose(CMC) and seemed to has transglycosylation activity simultaneously. Cellulase activity staining(zymogram) showed that the cellulase components of P. verruculosum were not aggregated in the medium. P. vrttuculosum mRNA was translated in vitro and translation product by the mRNA coding for CMCase IV was identified by immunoprecipitation.

섬유소 분해균인 P. verrculosum의 배양 여액으로부터 endo형 cellulase 인 CMCase IV를 정제하였다. CMCase IV는 13%의 탄수화물과 4.0의 pl값을 가진 산성, 당단백질이었다. CMCase IV의 SDSPAGE 상에서 분자량은 52 KDa이었으며, 효소 활성을 위한 최적 pH 및 온도는 5.0과 $50^{\circ}C$ 였다. CMCase IV를 CMC에 반응시 대부분 glucose와 cellobiose가 생산되었으며, 또한 동시에 transglycosylation 작용을 함께 갖는 것으로 사료되었다. Cellulase 활성 염색법(zymogram)을 통해서 P. verruculosum의 cellulase component가 배지 내에서 aggregation 되어있지 않음을 알 수 있었다. P. verruculosum mRNA의 in vitro 번역을 통하여 CMCase IV를 coding하는 번역산물이 동정 되었다.