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40.從雞糞堆肥篩選產生胞外蛋白酶之微生物

廖仁寶⁽¹⁾　蕭庭訓⁽¹⁾　楊琇雅⁽¹⁾　郭猛德⁽¹⁾　張秀鑾⁽²⁾　吳明哲⁽¹⁾　李佳音⁽³⁾

⁽¹⁾行政院農業委員會畜產試驗所

⁽²⁾國立屏東科技大學畜產系

⁽³⁾國立台灣大學農業化學系

本研究總共使用約3.5立方公尺之鷄糞於發酵槽中製作堆肥，並持續記錄堆肥分別為30、50及70公分深之溫度變化。於堆肥50公分處深測得發酵最高溫度約66℃，並進行採樣。將樣品分別保存於4℃冷藏櫃與凍存於-70℃冷凍櫃中。將冷藏之堆肥樣品溶於滅菌水後，再分別塗抹於胰蛋白腖大豆洋菜培養基與含5%脫脂奶粉之腖蛋白大豆洋菜培養基上，在50℃培養下胰蛋白腖大豆洋菜培養基上能長出眾多不同之菌落，並可在含5%脫脂奶粉之腖蛋白大豆洋菜培養基挑選到產生胞外蛋白酶之菌株共八株。由16S rDNA序列分析得知，此八株菌除一株為產鹼桿菌屬外，其他皆為桿菌屬。當這些分離株與商業化或專利菌株比較其蛋白酶活性時，有些分離株的活性比較高。因此，有些分離株具有進一步研究的潛力。

關鍵語：雞糞堆肥、胞外蛋白酶、發酵槽

SCREENING FOR EXTRACELLULAR PROTEASE PRODUCING
MICROORGANISMS FROM CHICKEN MANURE COMPOST

R. B. Liaw⁽¹⁾, T. S. Hsiao⁽¹⁾, H. Y. Yang, M. T. Kuo⁽¹⁾, H. L. Chang⁽²⁾, M. C. Wu⁽¹⁾ and C. Y. Lee⁽³⁾

⁽¹⁾ Livestock Research Institute, Council of Agriculture, Executive Yuan

⁽²⁾ Department of Animal Science, National Pingtung University of Science and Technology

⁽³⁾ Department of Agricultural Chemistry, National Taiwan University

A total of 3.5 cubic meter of chicken manure was used for composting in fermentor in this study. The temperature profiles of depth of 30, 50 and 70 cm, respectively, under the compost were continuously recorded. During the composting, the highest temperature under the 50 cm depth of compost was 66 ℃ and then the compost was sampled in the region. The samples were preserved in 4 ℃ refrigerator and -70 ℃ freezer, respectively. The sample in the refrigerator was weighed 0.5 gram to be dissolved in 20 ml of autoclaved water and 100 μl of the suspension was spread on TSA and TSA-5 % skim milk plates, respectively. There were many different colonies formed on TSA plate at 50 ℃ of incubation. Eight isolates capable of secreting extracellular proteases were obtained from TSA-5 % skim milk plates. Identifying the eight isolates by 16S rDNA sequencing, only one isolate belonged to Alcaligenes genus, and the others Bacillus genus. While comparing the protease activity, some of the isolates were superior to the commercialized and patented strains. Therefore, there was potential for studying the isolates in the future.

Key Words: Chicken manure compost, Extracellular protease, Fermentor