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Assisting biorecycling of bioorganic wastes by enzymatic approaches

通过酶法协助生物有机废物的生物回收

基本信息

批准号：
12480160
负责人：
NAKAYAMA Toru
金额：
$ 1.02万
依托单位：
TOHOKU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12480160/
关键词：
Enzyme Bioorganic wastes Compost Recycle Collagen Gelatin Garbage wastes lipids 耐熱性

项目摘要

We isolated an acidophilic thermophile belonging to the genus Bacillus, strain NTAP-1, which secreted a thermostable collagenolytic activity into the culture medium. The collagenolytic activity exhibited an optimum ph for Azocoll hydrolysis of ph 3.9 and was not completely inhibited by 10 Mm ethylenediaminetetraacetic acid (residual activity, 63 % ), suggesting that Bacillus NTAP-1 produces a novel acid proteinase with highest activity for collagen. The collagenolytic activity was thermostable; more than 80 % of the original activity was retained after incubation of the culture supernatant at ph 4.0 and 60 ℃ for 4h. We also isolated a Kpolytic bacterium, strain No. 6, from Siberian tundra soil. It was a Gram-negative coccoid rod capable of growing at 4 ℃ but not at 37 ℃ and was identified as a psychrotrophic strain of the genus Acinetobacter. Strain No.6 extracellularly produced a lipolytic enzyme that efficiently hydrolyzed triglycerides such as soybean oil during bacterial growth even at 4 ℃ ; it degraded 60 % of added soybean oil (initial concentration, 1 % w/v) after cultivation in LB medium at 4 ℃ for 7d. Thus, the bacterium is potentially applicable to in-situ bioremediation or bioaugumentation of fat-contaminated cold environments. We partially purified the lipolytic enzyme from the culture filtrate by acetone fractionation and characterized it.

我们分离出属于芽孢杆菌属的嗜酸耐热菌 NTAP-1 菌株，它向培养基中分泌热稳定的胶原蛋白分解活性。胶原蛋白水解活性表现出 Azocoll 水解的最佳 pH 值为 3.9，并且不被 10 Mm 乙二胺四乙酸完全抑制（残余活性，63%），表明芽孢杆菌 NTAP-1 产生了一种对胶原蛋白具有最高活性的新型酸性蛋白酶。溶胶原活性是热稳定的；培养上清液在pH 4.0、60℃培养4h后仍保留80%以上的原始活性。我们还从西伯利亚苔原土壤中分离出了一种 Kpolytic 细菌，即 6 号菌株。它是一种革兰氏阴性球状杆状体，能在4℃下生长，但不能在37℃下生长，经鉴定为不动杆菌属耐冷菌株。 6号菌株在细胞外产生一种脂肪分解酶，即使在4℃的细菌生长过程中也能有效水解甘油三酯，例如大豆油；在LB培养基中4℃培养7d后，其降解了60%添加的大豆油（初始浓度，1%w/v）。因此，该细菌可能适用于脂肪污染的寒冷环境的原位生物修复或生物强化。我们通过丙酮分级分离从培养物滤液中部分纯化了脂肪分解酶并对其进行了表征。