丙酸菌HZ-P-35的分离鉴定及其生成共轭亚油酸的研究

从奶牛青贮饲料中分离筛选出1株丙酸菌菌株HZ-P-35,液相色谱检测证明该微生物能从葡萄糖生产丙酸、乙酸和虎珀酸。通过平板菌落形态、液体培养、透射电镜等形态学手段观察、生理生化实验鉴定以及16SrRNA序列分析,初步确定所得菌株HZ-P-35为1株费氏丙酸杆菌变种,命名为Propionibacterium frudenreichiivar LL。细胞发酵转化实验结果表明,菌株HZ-P-35在MRS培养基中具有转化亚油酸生成共轭亚油酸的能力,气相色谱测定表明共轭亚油酸的质量浓度达24.37μg/mL。     

丙酸高产菌株的选育及发酵培养基优化

通过环境压力筛选获得了一株耐受30 g/L丙酸的产酸丙酸杆菌菌株(Propionibacterium acidipropionici WY320),降低了发酵过程中丙酸的反馈抑制作用,采用正交实验设计优化了发酵培养基. 结果表明,发酵培养基最适的玉米浆、酵母膏和(NH4)2SO4浓度分别为60, 10和7.5 g/L. 该条件下,摇瓶培养耐酸菌株的发酵周期为240 h,丙酸产量为49.35 g/L,较优化前提高72.98%,产率为0.21 g/(L×h); 5 L发酵罐培养的发酵周期为168 h,丙酸产量达51.96 g/L,产率为0.31 g/(L×h),较摇瓶培养提高47.62%,优于文献报道水平.     

细胞膜分离费氏丙酸杆菌离位转化合成维生素B12

采用膜分离技术将费氏丙酸杆菌菌体细胞分离,离位合成转化维生素B12,考察了不同类型及孔径的膜对发酵液的处理能力及对菌体细胞活性的损伤情况,分析了不同浓缩条件下维生素B12的转化效果. 结果表明,卷式膜比陶瓷膜更适用于发酵液中菌体细胞的分离,用0.22 mm卷式膜处理时,细胞的生物活性几乎不受损伤,存活率达99.5%,将发酵液浓缩4倍,中间体Abo-cbi转化率达78.8%,维生素B12产量为53.9 mg/L.     

基于离子交换层析技术提取发酵液中的丙酸

用7种不同性能的阴离子交换树脂吸附发酵液中丙酸,考察了发酵液酸化方式及pH值对吸附量的影响,分析了不同浓度H2SO4的解吸情况,比较了多柱串联组合层析提高丙酸收率的可行性. 结果表明,大孔弱碱性丙烯酸系阴离子交换树脂ZGD630对发酵液中的丙酸吸附量最高,采用H型阳离子交换树脂D001酸化工艺,当pH值由6.5降至3时,ZGD630树脂对发酵液中丙酸的静态吸附量从54.34 mg/g提高至110 mg/g,动态吸附量达152.5 mg/g. 用4 mol/L H2SO4进行解吸,丙酸浓度最高达81.8 g/L,为初始发酵液的2.12倍. 采用3柱串联组合层析时丙酸收率达62.15%,较单柱层析显著提高.     

Generation of volatile fatty acids by axillary bacteria

It is generally accepted that short-chain (C(2)-C(5)) volatile fatty acids (VFAs) are among the causal molecules of axillary malodour. It is also widely acknowledged that malodour generation is attributable to the biotransformation of odourless natural secretions, into volatile odorous products, by axillary bacteria. However, little information is available on the biochemical origins of VFAs on axillary skin. In these studies, assay systems were developed to investigate the generation of VFAs from substrates readily available to the bacteria resident on axillary skin. Propionibacteria and staphylococci were shown to ferment glycerol and lactic acid to the short-chain (C(2)-C(3)) VFAs, acetic and propionic acid. Furthermore, staphylococci are capable of converting branched aliphatic amino acids, such as leucine, to highly odorous short-chain (C(4)-C(5)) methyl-branched VFAs, such as isovaleric acid, which are traditionally associated with the acidic note of axillary malodour. However, in vitro kinetic data indicates that these pathways contribute less to axillary VFA levels, than fatty acid biotransformations by a recently defined sub-group of the Corynebacterium genus, corynebacteria (A). The results of these studies provide new understanding on the biochemical origins of VFA-based axillary malodour which, in turn, should lead to the development of novel deodorant systems.     

新型混菌发酵制备丙酸工艺及其优化

以产酸丙酸杆菌和酵母菌为菌种,进行混菌发酵制备丙酸的工艺研究,并探索最优工艺条件。通过单因素实验和正交优化实验确定了最优发酵工艺条件为:酿酒酵母为辅助菌,接种比例为Pa∶Sc=2.5∶1,接种间隔时间为8h,发酵温度为30℃,以NaOH(2mol/L)溶液为中和剂,采用自动流加方式,发酵时间为180h。优化后的混菌发酵工艺能有效提高丙酸产量,丙酸产量达24.16g/L,与纯菌发酵相比提高46.34%。