MEMETIMIN Henipigul, METQURBAN Emerjan, BUGHDASH Ayan, et al. Isolation of an α=amylase producing Bacillus and optimization of its fermentation conditions[J]. Acta Scientiarum Naturalium Universitatis SunYatseni, 2017,56(4):126-132.
MEMETIMIN Henipigul, METQURBAN Emerjan, BUGHDASH Ayan, et al. Isolation of an α=amylase producing Bacillus and optimization of its fermentation conditions[J]. Acta Scientiarum Naturalium Universitatis SunYatseni, 2017,56(4):126-132.DOI:
以筛选与鉴定尉犁县黑湖产淀粉酶细菌,并对筛选出的细菌进行产酶条件优化为研究目的,从43株细菌中筛选出淀粉酶高产菌株 HM-22,并对其进行了菌株形态学鉴定、16S r DNA 分子鉴定以及产酶条件优化。从尉犁县黑湖采样的30个水、土和泥样样品中分离筛选43株细菌,从中筛选出14株产淀粉酶的菌株,采用Yoo改良法对产透明圈较大的菌株进行酶活力测定,从中筛选出了一株产酶活性较高的菌株HM-22。经革兰氏染色、菌落形态观察、生理生化检测和16S r DNA 序列比对鉴定该菌与Bacillus tequilensis 的相似性为99.8%,属于芽孢杆菌属。对菌株HM-22产酶条件进行初步优化确定其产酶的最佳条件是:该菌产淀粉酶最适温度为40 ℃,最适碳源为可溶性淀粉,最适氮源为牛肉膏,最适反应pH为6.0,最适产酶培养时间为24 h。优化后菌株HM-22的酶活力从最初的的酶活力122.45 U/mL达到147.53 U/mL,酶活力提高了17%。该研究为从新疆盐湖细菌中筛选淀粉酶活力较高的菌种资源及应用提供了理论依据和参考。
Abstract
The purpose of the study is to screen and identify amylase producing bacteria in the Black Lake in Yuli county
and to optimize the conditions of enzyme production. A high amylase producing strain HM-22 was screened from 43 strains of bacteria
has carried out morphological identification
molecular identification of 16S rDNA and optimization of enzyme production conditions were carried out. 43 strains of bacteria were isolated from 30 water
soil and mud samples in the Black Lake in Yuli County
and 14 strains producing amylase were screened out. The enzyme activity was determined by Yoo method
a strain with high enzyme activity was obtained and named as HM-22. By gram staining
colony morphology observation
physiological and biochemical tests
16S rDNA sequence alignment and the similarity of Bacillus tequilensis was 99.8%
belonging to the genus Bacillus. The optimization of enzyme production conditions of strain HM-22 was determined and the best condition for producing the enzyme is: the optimum temperature of amylase production was 40 ℃
the optimal carbon source is soluble starch
the optimal nitrogen source was beef extract
the optimum reaction pH was 6.0
the optimum culture time was 24 h. After optimization
the enzyme activity of strain HM-22 reached 147.53 U/mL from the initial activity of 122.45 U/mL
enzyme activity increased by 17%. The study provides a theoretical basis and reference for the screening of strains with high amylase activity in Xinjiang salt lake bacteria