生物加工过程
生物加工過程
생물가공과정
CHINESE JOURNAL OF BIOPROCESS ENGINEERING
2014年
4期
87-93
,共7页
何剑%雍晓雨%周俊%王舒雅%陈怡露%郑涛
何劍%雍曉雨%週俊%王舒雅%陳怡露%鄭濤
하검%옹효우%주준%왕서아%진이로%정도
解淀粉芽胞杆菌%鉴定%分离筛选%γ 多聚谷氨酸
解澱粉芽胞桿菌%鑒定%分離篩選%γ 多聚穀氨痠
해정분아포간균%감정%분리사선%γ 다취곡안산
Bacillus amyloliquefaciens%identification%isolation%γ-polyglutamic acid(γ-PGA)
从菜园土壤中取样，在含有谷氨酸的筛选培养基上采用梯度稀释涂布、平板划线的方法，以菌落/菌液黏稠度为指示，分离筛选生产γ多聚谷氨酸的菌株。利用氨基酸分析仪测定提取纯化后的γ多聚谷氨酸的产量，并通过形态学、生理生化特征以及16S rDNA基因序列分析鉴定该菌株，并对其合成γ多聚谷氨酸的功能基因进行PCR扩增。结果表明：筛选到1株产γ多聚谷氨酸的细菌C1，其液体摇瓶发酵产量为18?4 g/L，相对分子质量为1?8×106；该菌株为革兰氏阳性，菌落黏稠、菌体呈杆状、产芽胞、且形成荚膜；主要生理生化特点为能利用葡萄糖和蔗糖发酵，水解淀粉，H2 O2酶阳性，产吲哚等；经16S rDNA鉴定与Bacillus amyloliquefaciens ATCC 23350同源性为100％，故命名为Bacillus amyloliquefaciens C1，且拥有γ多聚谷氨酸合成的相关基因pgsA、pgsB和pgsC。
從菜園土壤中取樣，在含有穀氨痠的篩選培養基上採用梯度稀釋塗佈、平闆劃線的方法，以菌落/菌液黏稠度為指示，分離篩選生產γ多聚穀氨痠的菌株。利用氨基痠分析儀測定提取純化後的γ多聚穀氨痠的產量，併通過形態學、生理生化特徵以及16S rDNA基因序列分析鑒定該菌株，併對其閤成γ多聚穀氨痠的功能基因進行PCR擴增。結果錶明：篩選到1株產γ多聚穀氨痠的細菌C1，其液體搖瓶髮酵產量為18?4 g/L，相對分子質量為1?8×106；該菌株為革蘭氏暘性，菌落黏稠、菌體呈桿狀、產芽胞、且形成莢膜；主要生理生化特點為能利用葡萄糖和蔗糖髮酵，水解澱粉，H2 O2酶暘性，產吲哚等；經16S rDNA鑒定與Bacillus amyloliquefaciens ATCC 23350同源性為100％，故命名為Bacillus amyloliquefaciens C1，且擁有γ多聚穀氨痠閤成的相關基因pgsA、pgsB和pgsC。
종채완토양중취양，재함유곡안산적사선배양기상채용제도희석도포、평판화선적방법，이균락/균액점주도위지시，분리사선생산γ다취곡안산적균주。이용안기산분석의측정제취순화후적γ다취곡안산적산량，병통과형태학、생리생화특정이급16S rDNA기인서렬분석감정해균주，병대기합성γ다취곡안산적공능기인진행PCR확증。결과표명：사선도1주산γ다취곡안산적세균C1，기액체요병발효산량위18?4 g/L，상대분자질량위1?8×106；해균주위혁란씨양성，균락점주、균체정간상、산아포、차형성협막；주요생리생화특점위능이용포도당화자당발효，수해정분，H2 O2매양성，산신타등；경16S rDNA감정여Bacillus amyloliquefaciens ATCC 23350동원성위100％，고명명위Bacillus amyloliquefaciens C1，차옹유γ다취곡안산합성적상관기인pgsA、pgsB화pgsC。
Soil sample was collected from farmland. The strains were screened by the methods of gradient dilution and plate streaking on the selected medium containing glutamic acid and under the indication of the viscosity of their fermentation broth. The γ-polyglutamic acid production of the strains was obtained by analyzing the glutamic acid content of the hydrolysate of γ-polyglutamic acid using amino acid analyser. Then, the isolated strain was identified according to the morphological features, physiological and biochemical characteristics, as well as phylogenetic analysis of 16S rDNA sequences. Finally, the pgsA, pgsB, and pgsC gene, related to the γ-polyglutamic acid synthesis were amplified by PCR. A Bacillus, called the C1, was isolated with aγ-polyglutamic acid production of 18?4 g/L in flask fermentation culture. The molecular mass ofγ-polyglutamic acid produced by strain C1 was 1?8×106 detected by GPC. C1 was a gram-positive bacilli, which fermented both glucose and sucrose, hydrolysed starch, produced indole, and shared 100% sequence identity of its 16S rDNA with Bacillus amyloliquefaciens ATCC 23350, thus it was called the Bacillus amyloliquefaciens C1. It was testified to harboring pgsA, pgsB, and pgsC genes.
