混菌发酵绿茶产脂肪酶及其对酯型儿茶素的水解作用分析

微生物的发酵作用导致黑茶加工过程中儿茶素的生物转化和总茶多酚含量的下降,这一转化过程的代谢途径和调控机制尚不清楚。通过添加冠突曲霉（Aspergillus cristatum）菌株PE-1和黑曲霉（A. niger）菌株PE-4混合发酵绿茶,采用鉴别培养基平板检测、酶活性测定和酶蛋白分离纯化,研究脂肪酶活性及其对酯型儿茶素的水解作用。两个菌株单菌发酵和混合菌种发酵绿茶条件下均可以检测到脂肪酶活性,混菌发酵酶活力高于两个菌株单菌发酵的酶活力。混菌发酵条件下获得一个分子量约为37βkDa的脂肪酶,对EGCG和ECG有不同的生物转化作用。UPLC检测结果表明,该脂肪酶对EGCG有较好的底物选择性,EGCG的水解率为94.46%,EGC的产率为63.33%;ECG的水解率为15.45%,EC的产率为4.15%。脂肪酶对EGCG和ECG的生物转化将为儿茶素代谢途径的研究和单体儿茶素的生产起到促进作用。     

微生物转化人参皂苷Rb1为稀有皂苷F2

从橙汁中分离到的菌株CZ2能使人参皂苷Rb1转化为稀有皂苷F2和gypenoside-XVII.最佳转化培养基含酵母提取粉1 g/L、氯化铵0.5 g/L、磷酸氢二钾1 g/L、磷酸二氢钾0.5 g/L、硫酸镁0.25 g/L,pH值为4.0.菌株CZ对人参皂苷Rb1的生物转化机理为Rb1→Gpy-XVII→F2.     

鞘氨醇单胞菌2-F2将人参主皂苷Re转化为人参稀有皂苷Rh1

从人参根系土壤中分离的菌株2-F2将人参主皂苷Re转化为稀有皂苷Rh1,其转化机理为Re→Rg1→Rh1.进行16S rRNA基因序列的系统发育分析,菌株2-F2与Sphingomonas asaccharolytica IFO15499T在同一分支上,同源性为99.5%.该菌株属于鞘氨醇单胞菌属.     

3β,5α,6β-三羟基-16-孕甾烯酮的生物合成及晶体结构分析

利用绿色木霉对5,6α-环氧-3β-羟基-5α-孕甾-16-稀-20-酮进行生物转化得到3β,5α,6β-三羟基孕甾-16-烯-20-酮和3β,5α,6β,15β-四羟基孕甾-16-稀-20-酮。利用IR、MS和2D-NMR实验技术等波谱方法对生物转化产物的结构进行了详细研究,运用X-射线单晶衍射方法确定3β,5α,6β-三羟基孕甾-16-烯-20-酮的空间构型,并进行了晶体结构分析。     

一株脱氧雪腐镰刀菌烯醇转化菌的分离、鉴定及其转化效果研究

以脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)为唯一碳源,通过富集培养从土壤中分离到一株可以转化DON的真菌,命名为NJA-1.用引物Bt2a、Bt2b扩增了该菌的β-tubulin基因序列,GenBank的登录号为DQ902579.通过基因序列分析和形态学观察最终鉴定该菌株为塔宾曲霉(Aspergillus tubingensis).该菌与塔宾曲霉AY820009的同源性为99%.在pH 60、30 ℃、14 d内其对DON的平均转化率为944%.HPLC-MS分析结果表明,DON转化产物的相对分子质量为3144,比DON的相对分子质量(2963)增加了181.该菌在无机盐培养基中对DON的最高耐受浓度为40 mg·L-1.提示:该菌可用于DON的生物转化.     

Characterisation of the substrate specificity of the nitrile hydrolyzing system of the acidotolerant black yeast Exophiala oligosperma R1

The ‘black yeast’ Exophiala oligosperma R1 can utilise various organic nitriles under acidic conditions as nitrogen sources. The induction of a phenylacetonitrile converting activity was optimised by growing the strain in the presence of different nitriles and /or complex or inorganic nitrogen sources. The highest nitrile hydrolysing activity was observed with cells grown with 2-cyanopyridine and NaNO₃. The cells metabolised the inducer and grew with 2-cyanopyridine as sole source of nitrogen. Cell extracts converted various (substituted) benzonitriles and phenylacetonitriles. They usually converted the isomers carrying a substituent in the meta-position with higher relative activities than the corresponding para- or ortho-substituted isomers. Aliphatic substrates such as acrylonitrile and 2-hydroxy-3-butenenitrile were also hydrolysed. The highest specific activity was detected with 4-cyanopyridine. Most nitriles were almost exclusively converted to the corresponding acids and no or only low amounts of the corresponding amides were formed. The cells hydrolysed amides only with extremely low activities. It was therefore concluded that the cells harboured a nitrilase activity. The specific activities of whole cells and cell extracts were compared for different nitriles and evidence obtained for limitation in the substrate-uptake by whole cells. The conversion of 2-hydroxy-3-butenenitrile to 2-hydroxy-3-butenoic acid at pH 4 demonstrated the unique ability of cells of E. oligosperma R1 to hydrolyse aliphatic α-hydroxynitriles under acidic conditions. The organism could grow with phenylacetonitrile as sole source of carbon, energy and nitrogen. The degradation of phenylacetonitrile presumably proceeds via phenylacetic acid, 2-hydroxyphenylacetic acid, 2,5-dihydroxyphenylacetic acid (homogentisate), maleylacetoacetate and fumarylacetoacetate.     

Biotransformation of tuna waste by co-fermentation into an aquafeed ingredient

Dried skipjack tuna ( Katsuwonus pelamis) waste (red meat, gills, viscera, fins, etc.) were mixed with 25% wheat flour and inoculated with a starter culture of Lactobacillus plantarum National Collection of Industrial Microorganisms (NCIM) 2912 (10⁸–10⁹ cells mL⁻¹) and Bacillus licheniformis MTCC 6824 (10⁷–10⁸ cells mL⁻¹). Changes in the nutritional quality (crude protein, crude fat, crude ash, crude fibre and nitrogen-free extract and aminoacids) were monitored during a fermentation period of 14 days. The proximate analysis showed significant changes in the composition of L. plantarum -fermented tuna (LPFT) and B. licheniformis -fermented tuna (BLFT) from the unfermented raw materials. Fermentation of tuna waste has resulted in a significant ( P <0.05) increase in the protein content of tuna waste between days 6 and 12. All the amino acid contents in BLFT increased during fermentation, whereas, in LPFT the levels of serine, histidine, tyrosine, methionine, cystine and phenylalanine contents were decreased. A marginal increase in calcium and phosphorus levels was recorded in the fermented products. The results of the study suggest that LPFT or BLFT can be used as a novel aquafeed ingredient for different fish species.     

黑茶发酵优势菌对茶多酚的生物转化研究

利用从茯砖茶中分离出的冠突散囊菌和从普洱茶中分离出的黑曲霉、根霉分别接种于以茶多酚作为唯一碳源的培养基中,进行单一菌株发酵,对发酵过程茶多酚类化合物的变化规律进行分析.结果表明,发酵期内,随着发酵时间延长,各发酵液的茶多酚总量显著降低,冠突散囊菌、黑曲霉、根霉发酵液中多酚分别降低38.9％、85.5％和92.1％;黄酮类总量在黑曲霉、根霉的作用下先下降后上升,冠突散囊菌则波动上升;各处理儿茶素总量均显著下降,其中酯型儿茶素含量直线下降,非酯型儿茶素和没食子酸的含量均先增加后减少.在黑曲霉、根霉作用下,茶黄素、茶红素含量显著减少,茶褐素含量增加.冠突散囊菌发酵液中茶黄素含量减少,茶红素含量增加,茶褐素含量基本不变.基于不同优势菌对茶多酚的转化在质和量上均有差异,有必要对其转化产物进行深入研究.     

自然铜的生物转化研究

利用Acidithiobacillus ferrooxidans ATCC23270(A.ferrooxidans ATCC23270)对自然铜进行生物转化,考察矿浆浓度,微生物接种量,系统初始pH,浸出时间以及温度对自然铜转化的影响。利用单因素法对A.ferrooxidans转化自然铜进行条件优化。结果表明,自然铜最佳浸出工艺为矿浆浓度0.5%、系统初始pH值1.75、接种量10%、浸出时间15 d以及环境温度35℃,浸出率可达65.32%。初步获得了A.ferrooxidans转化自然铜的条件,为中药的炮制提供新思路。     

Hydroxysteroid Dehydrogenases in Bovine and Porcine Granulosa Cells Convert Zearalenone into its Hydroxylated Metabolites α-Zearalenol and β-Zearalenol

The enzymes 3α- and 3β-hydroxysteroid dehydrogenase (3α- and 3β-HSD) play a pivotal role in synthesis of various steroid hormones including oestradiol and testosterone. The structure of the mycotoxin zearalenone resembles many characteristics of steroids and binds to oestrogen receptors as an agonist. Consequently, it is suggested that zearalenone is also a substrate for 3α-HSD and 3β-HSD. 3α-HSD and 3β-HSD isoforms are expressed in the liver and kidney but also in many steroidogenic tissues. It was the aim of the present study to demonstrate the presence of these enzymes in granulosa cells, which were obtained from bovine and porcine ovaries, and to investigate whether zearalenone is a substrate for these enzymes. The results show a species-specific expression pattern in the granulosa cells of both species. Moreover, it was demonstrated that zearalenone when added to the culture medium, is converted into α-zearalenol and β-zearalenol. Corresponding to the apparent expression profile, in porcine granulosa cells predominantly α-zearalenol was formed, whereas bovine granulosa cells preferentially converted zearalenone into β-zearalenol. This is the first report demonstrating the extrahepatic biotransformation of zearalenone in target tissues.