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Characteristics of α-Amylase and Protease Produced from Bacillus amyloliquefacies CNL-90 Isolated from Malt Grain

맥아에서 분리한 Bacillus amyloliquefacies CNL-90이 생산하는 α-amylase와 Protease의 특성

  • Bae, Hyoung-Churl (Lab. of Milk Food Biochemistry and Biotechnology College of Agriculture and Life Sciences Chungnam National University) ;
  • Choi, Seong-Hyun (Research Institute, KG Si-Tech.) ;
  • Na, Seuk-Han (Chung Mi Bio Inc.) ;
  • Nam, Myoung-Soo (Lab. of Milk Food Biochemistry and Biotechnology College of Agriculture and Life Sciences Chungnam National University)
  • 배형철 (충남대학교 농업생명과학대학 동물바이오시스템과학과) ;
  • 최성현 ;
  • 나석한 (청미바이오) ;
  • 남명수 (충남대학교 농업생명과학대학 동물바이오시스템과학과)
  • Received : 2012.01.30
  • Accepted : 2012.04.24
  • Published : 2012.04.30

Abstract

A bacterium, identified as $Bacillus$ $amyloliquefacies$, CNL-90 using 16S rDNA analysis, was isolated from malt grain. The optimal activities of its ${\alpha}$-amylase and protease were observed at pH 6 and $60^{\circ}C$, and at pH 6 and $50^{\circ}C$, respectively although their activities remained stable at pH 7 and $40^{\circ}C$for ${\alpha}$-amylase and at pH 7 and $50^{\circ}C$ for protease. After solid-state fermentation of $B.$ $amyloliquefacies$, CNL-90 on wheat bran for 72hr or 144hr, the ${\alpha}$-amylase and protease activities were 170,000 and 290,000 units/kg, and 290,000 and 310,000 units/kg, respectively. The viable bacterial cell counts were $1.5{\times}10^9$ CFU/g and $2.2{\times}10^9$ CFU/g at 72hr and 144hr of the solid-state fermentation, respectively. A feeding trial with a total of 127 piglets was also conducted. The animals were divided into two groups: an experimental group fed with the fermented product (63 piglets) and a control group (64 piglets). The growth rate of the experimental group was 6.66% higher than that of the control group (P<0.05). The results of this study indicate that the ${\alpha}$-amylase and protease from $B.$ $amyloliquefacies$, CNL-90 can be used for industrial applications due to their activity in production of carbohydrate hydrolysates.

Bacillus 균주가 생산하는 효소를 식품과 동물사료 첨가제로 이용하기 위해서 맥아로부터 단백질 분해능력과 전분 분해능력이 우수한 균주를 분리 동정하여 $Bacillus$ $amyloliquefaciens$ CNL-90으로 명명하였다. 분리된 $B.$ $amyloliquefaciens$ CNL-90이 생산하는 ${\alpha}$-amylase의 안정성은 pH는 7, 온도는 $40^{\circ}C$, protease의 경우는 pH가 7, 온도는 $50^{\circ}C$에서 안정했다. $B.$ $amyloliquefaciens$ CNL-90을 밀기울에 접종하여 고체 배양한 결과 ${\alpha}$-amylases의 효소활성은 6일 배양 후 290,000 unit/kg이었고, protease의 효소활성은 310,000 unit/kg으로 나타났다. 밀기울에 고체 배양한 생균수는 배양 6일 후에는 $2.2{\times}10^9$ CFU/g으로 높았다. 사료 요구율 개선은 $B.$ $amyloliquefaciens$ CNL-90 배양액 0.2% 첨가구가 대조구에 비해서 일당 증체량은 6.66% 높았고, 사료 요구율은 0.05% 효과가 있었다. 이러한 결과는 $B.$ $amyloliquefaciens$ CNL-90이 생산하는 단백질 분해효소와 전분 분해효소는 식품산업 및 가축사료 첨가제 산업에 응용할 수 있는 가능성을 확인하였다.

Keywords

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