摘要
【目的】对塔吉克斯坦土壤细菌产功能酶菌株筛选及产淀粉酶的酶学特性的初步研究。【方法】采用透明圈法对从塔吉克斯坦土壤细菌产几种重要的工业用功能酶筛选。通过形态学观察、16S r DNA和gyr B基因序列测定对一株产高温淀粉酶的菌株进行鉴定,研究其淀粉酶的酶学性质。【结果】从塔吉克斯坦土壤中共分离细菌菌株110个,其中有产淀粉酶菌株70个、产蛋白酶菌株56个、产纤维素酶菌株37个、产脂肪酶菌株16个,占总菌株数比例依次为63.6%、50.9%、33.6%、14.5%。对筛选出产淀粉酶活性较高的T-17菌株的粗酶学性质测定表明,该淀粉酶具有高温、中性、耐盐淀粉酶的特性,最适催化温度是70℃,30~70℃维持较高酶活力,最适催化p H值为7.0,p H值在较稳定6.0~8.0,有较强的耐盐特性,在4 mol/L的Na Cl浓度下仍能保持较高的酶活性,Ca2+和K+对酶具有较强的激活作用。经形态观察和分子生物学方法将T-17鉴定为巨大芽孢杆菌(Bacillus malacitensis)。【结论】塔吉克斯坦土壤微生物产功能酶多样性丰富,筛选获得的高温淀粉酶具有一定的应用潜力。
【Objective】The strains producing functional enzyme were isolated from soil in Tajikistan and their characteristics of amylase were researched.【Method】Soil bacteria producing several kinds of important industrial enzyme were screened by transparent circle method in Tajikistan. A strain producing high temperature amylase was identified through morphological observation and sequencing of 16 S r DNA,gyr B gene.【Result】110 soil bacteria were isolated from the soil in Tajikistan,among which 70 strains produced amylase,56 strains produced protease,37 strains produced cellulose and 16 strains produced lipase,and the proportion of the total number of strains were 63. 6 %,50. 9%,33. 6% and 14. 5% respectively. Coarse enzymology properties determination of T- 17 strain producing high amylase activity showed that the amylase had the characteristics of high temperature,neutral and salt tolerance,and the optimal catalytic temperature was 70 ℃,which still had higher enzyme activity at 30 ~ 70℃. The optimal catalytic p H was 7. 0 and a relatively stable in p H6. 0 ~ 8. 0 with stronger salt- tolerant characteristics. When the concentration of Na Cl was 4 mol / L,it could still keep high enzyme activity. Ca2 +and K+had apparent activation to the enzyme. The strain of T- 17 was identified as Bacillus anthracoides through morphological observation and molecular method.【Conclusion】The soil microorganism producing functional enzyme was diverse,and the development and application of screening high temperature amylase had a certain potential in the future. This research work in this area provided the basis for a further development and utilization of the resources in the area.
出处
《新疆农业科学》
CAS
CSCD
北大核心
2015年第12期2294-2304,共11页
Xinjiang Agricultural Sciences
基金
国家国际科技合作计划项目(2010DFA92720-13)
新疆溢达奖学金资助~~
关键词
塔吉克斯坦
土壤细菌
功能酶
淀粉酶
酶活力
Tajikistan
soil bacteria
functional enzyme
amylase
enzyme activity
