CAJ | 학술논문

本研究从实验室保藏高效分泌能降解异甘露聚糖的甘露聚糖酶的细菌F1-5出发,采用菌株的个体和群体的形态观察、生理生化实验及16S rDNA系统发育分析手段进行鉴定,最终确定菌株F1-5为枯草芽孢杆菌(Bacillus subtilis).Genbank注册号为FJ392725.通过单因素试验和正交优化试验,确定枯草芽孢杆菌F1-5的最佳发酵产酶条件.酵母细胞壁碳源浓度为4.8%,氮源浓度为0.5%,培养基初始pH6.0,接种量为6%,装液量为60ml/250ml三角瓶,转速为180r/min,培养温度为35℃,在此发酵条件下发酵48h,枯草芽饱杆菌F1-5产生的甘露聚糖酶活力达330U/ml,是优化前的3.4倍.发酵罐扩大实验,甘露聚糖酶活力可达582U/ml.
본연구종실험실보장고효분비능강해이감로취당적감로취당매적세균F1-5출발,채용균주적개체화군체적형태관찰、생리생화실험급16S rDNA계통발육분석수단진행감정,최종학정균주F1-5위고초아포간균(Bacillus subtilis).Genbank주책호위FJ392725.통과단인소시험화정교우화시험,학정고초아포간균F1-5적최가발효산매조건.효모세포벽탄원농도위4.8%,담원농도위0.5%,배양기초시pH6.0,접충량위6%,장액량위60ml/250ml삼각병,전속위180r/min,배양온도위35℃,재차발효조건하발효48h,고초아포간균F1-5산생적감로취당매활력체330U/ml,시우화전적3.4배.발효관확대실험,감로취당매활력가체582U/ml.
A mannanase-producing bacterial strain F1-5 that isolated and preserved in our laboratory was identified as Bacillus subtilis (Genebank accession number FJ392725) according to the results of morphological observations, physiological and biochemical tests and 165 rDNA sequence homology analysis. The optimal fermentation conditions of F1-5 for producing mannanase were determined to be fermentation for 48 h at 35 ℃ with an inoculation size of 6% in a 250 ml flask with a shaking speed of 180 r/min containing 60 ml of culture medium consisting of 4.8% yeast cell wall as carbon source and 0.5% NH_4NO_3 as nitrogen source by one-factor-at-a-time and orthogonal array design methods. Under such conditions, the mannanase activity was up to 330 U/ml, which was 3.4 times higher than before the optimization. Furthermore, our 5 L fermentor experiments gave a mannanase activity as high as 582 U/ml. Therefore, these optimal conditions are reliable.