Enzyme Activity of Isolated Psychrotrophic Bacteria from Raw Milk of Different Regions on Season

The aim of this study was to investigate the effect of season and location on activities of enzyme produced by psychrotrophic bacteria isolated from raw milk located in Kyunggi region of South Korea. Agar diffusion and colorimetric methods were used for the lipase and protease activities of psychrotrophic bacteria. Intensities of dark blue and transparent ring around colony were compared for activity measurement. Nutrient agar with 1% skim milk added was employed for measuing protease activity. 14 strains of Arthrobacter russicus with lipase activity and 19 strains of Chryserobacterium shigense with protease activities were found to be present. It was found that Acinetobacter genomospecies 10 (match %: 99.90) isolated from B region in fall was the most lipolytic species, whereas Serratia liquefaciens (match %: 99.39) isolated from the same region in spring was the most proteolytic species. Growth curve of Acinetobacte and Serratia liquefaciens was a typical sigmoidal form. Lipase activity increased with incubation time, but its activity began to drop at stationary to motality phase. Optimum condition for incubation time, pH and temperature for extracellular lipase from Acinetobacter genomospecies 10 (match %: 99.90) was 12 h, 8.5, and $45^{\circ}C$, respectively. Extracellular protease from Serratia liquefaciens (match %:99.39) had the same optimum incubation time and pH as extracellular lipase, but optimum temperature was $35^{\circ}C$.

계절에 따라 여러 지역의 원유에서 분리된 내냉성 미생물의 효소 활성

지방분해효소 활성이 가장 높은 균은 가을철에 집유한 B 지역의 원유로부터 분리한 Acinetobacter genomospecies 10(match %: 99.90)이었으며, 단백질분해효소 활성이 가장 높은 균은 봄철에 B 지역의 원유로부터 분리한 Serratia liquefaciens(match %: 99.39)였다. Acinetobacter genomospecies 10(match %:99.90)는 전형적인 생장곡선의 형태인 시그모이드 곡선의 형태를 나타냈으며, 배양시간별 조효소 활성은 배양시간이 증가할수록 지방분해효소 활성이 증가하다가 정지기 후반부에서부터 사멸기에 도달하기까지는 배양시간이 길수록 활성이 감소하였다. pH에 따른 지방분해효소 활성은 pH 8.5에서 가장 높았고 온도는 $45^{\circ}C$에서 가장 높은 활성을 나타내었다. Serratia liquefaciens(match %: 99.39)의 생장곡선은 전형적인 시그모이드 형태를 나타내었고, 단백질분해효소 활성은 배양 12시간 후에 가장 높았고, 18시간 후에는 다시 감소하는 양상을 나타냈다. 단백질 분해효소의 pH 영향은 2.5에서 활성이 가장 낮았고 pH 8.5에서 가장 높은 활성을 나타내었다. 또한 반응온도에 의한 효소활성은 $35^{\circ}C$에서 가장 높았다.