Effect of Ozone on Aspergillus niger Causing Black Rot Disease in Onion

The effect of ozone on Aspergillus niger causing black rot disease in onion was studied in culture. Ozone induced the spore germination in all treatments and few spores showed rapid swelling, resulting in the production of 2-3 germ tubes per spore compared to control. Although all the ozone treated spores germinated, all of them did not produce uniform colony morphology. Some colonies which developed from ozone treated spores failed to produce spores and such colonies appeared as grey patches of mycelia without spores amidst surrounding black sporulating colonies. Further work is in progress to study the mechanism involved in formation of sterile mycelia by ozone.     

Purification and characterization of extracellular chitinase from a novel strain Aspergillus fumigatus CS-01

Twelve samples derived from different locations in south central area of China are treated by enrichment and spread-plate technique for initial screening. Seven chitinase-producing strains are isolated. The chitinase present in the culture supernatant of strain CS-01 possesses the maximum activity of 0.118 U/mL. Analysis of the morphological feature and the ITS rDNA sequence reveals that strain CS-01 belongs to Aspergillus fumigatus. Production of the chitinase is regulated by a inducible way and the maximum activity appears at 36 h in colloidal chitin culture. Purification of the chitinase is carried out by salting out, gel filtrate chromatography and anion exchange chromatography sequentially. Native-PAGE and SDS-PAGE indicate that the chitinase from A. fumigatus CS-01 is a monomer with the relative molecular mass estimated to be 4.50×10⁴. Its maximum activity appears at pH 5 and 55 °C. The chitinase is stable at pH 4.0–7.5 and below 45 °C. Foundation item: Projects(50621063, 50674101) supported by the National Natural Science Foundation of China     

米曲霉在牛乳中最适生长条件的研究

利用米曲霉对牛乳进行发酵,通过单因子及正交试验以菌丝体干重为指标对其在牛乳中的生长进行了研究．结果表明：接种量为4％,装液量为30mL,摇床转速为180r／min时菌丝体干重可达到最大．此外测得最大菌丝体干重时的蛋白质分解率可达35．6％．     

黑曲霉对含铅废水的生物吸附研究

研究黑曲霉真菌对污染废水中Pb^2＋的吸附．研究铅离子质量浓度、pH值、温度、灭菌与不灭菌以及重金属离子存在时对铅去除率的影响．结果表明：在实验条件下,Pb^2＋的最佳吸附质量分数是120mg／L,最适宜pH值为6．0,最佳温度为25℃．在20min取样时不灭菌对模拟废水中Pb^2＋的去除没有影响;20h时取样则由于环境中微生物与黑曲霉的竞争与协同作用使不灭菌的情况对铅的吸附明显好于灭菌的情况;干扰离子Hg^2＋、Cu^2＋、Zn^2＋的存在对铅的吸附无显著影响,并且在同一质量浓度情况下,这3种干扰离子的去除率大小为Zn^2＋〉Cu^2＋〉Hg^2＋．     

小麦中黄曲霉生长的预测模型

采用预测微生物学的方法研究了小麦中主要霉变微生物黄曲霉的生长规律,建立了小麦中黄曲霉生长的初级和二级预测模型.结果表明:在适温阶段黄曲霉的生长曲线呈典型的S形,用Gompertz模型拟合效果良好;当接近最低生长温度时,适合用线性回归方程拟合;小麦中黄曲霉生长的温度模型符合Rosso模型,并对模型的有效性进行了验证.根据预测模型,在相对湿度60%条件下,小麦中黄曲霉的最低生长温度是6.5℃,最高生长温度是52.7℃,最适生长温度是31.4℃.     

酱油曲菌的选择及酱油发酵条件

研究了日本酱油生产菌AspergillusoryzaeJP DQ 1,AspergillusoryzaeJP W和我国酱油生产菌AspergillusoryzaeAS3.951的酶系。结果表明:3种菌在浓酱油曲和白酱油曲培养基上均可产生α 淀粉酶、糖化酶、纤维素外切酶、果胶酶、酸性蛋白酶和中性蛋白酶;JP W是较好的白酱油生产菌,JP DQ 1是较好的浓酱油生产菌。还对酱油酵母的生理特性进行了部分研究,发现食盐浓度对酱油酵母生长影响较大,酱油发酵过程的盐浓度不宜超过3mol／L,低于2mol／L更佳。酱油酵母的较适生长pH为4.5～5.5。     

一株里氏木霉产纤维素酶的条件及酶系的优化

对一株里氏木霉突变株液体发酵产纤维素酶的培养条件进行了优化,确定培养基的最适碳源和氮源种类及添加量(g/L)分别为:小麦秸秆15,小麦麸皮5,尿素10～15,(NH4)2SO412,NH4NO35;最适培养条件为:接种种龄72h,接种孢子悬液浓度2×107个/mL,培养温度30～32℃,pH值5 0,培养时间144h,摇瓶转速180r/min,装液量75mL/250mL三角瓶.在培养里氏木霉的培养基中同时接种黑曲霉(二者孢子数比为60∶1)进行混合培养,β 葡萄糖苷酶的产量是对照试验的2 9倍,明显改善了纤维素酶系的比例,提高了它们之间的协同作用效果,在最适培养条件下,粗酶液的β 葡萄糖苷糖活和FPA酶活分别达到127 6U/mL和84 8U/mL.     

Enrichment of ω‐3 fatty acids in flax seed oil by alkaline lipase of Aspergillus fumigatus MTCC 9657

Flax seed oil obtained from the seeds of flax plant (Linum usitatissimum, L.) is an unexploited source which contains ω‐3 and ω‐6 fatty acids. Flax seed oil is hydrolysed with a novel alkaline lipase from Aspergillus fumigatus MTCC 9657 for the removal of unwanted fatty acids and enrichment of ω‐3 fatty acids. An appropriate balance of ω‐3 and ω‐6 polyunsaturated fatty acids and enzymatic enrichment of polyunsaturated fatty acids in diet promote nutrition and health. Fatty acid composition shows that flax seed oil contains about 26.80%, 13.5% and 25.45% of ω‐3 and ω‐6 fatty acids in triglyceride (TG), diglyceride (DG) and monoglyceride (MG), respectively. After 8 h of hydrolysis, ω‐3 content was increased to 39% in TG, showing that unwanted saturated fatty acids are removed. ω‐6 content of triglycerides in flax seed oil also showed 54.76% increase after 8 h of hydrolysis. An enzymatic method of hydrolysis by fungal lipase was developed by this study and achieved to concentrate the essential fatty acids linoleic acid (LA) and α‐linoleic acids (ALA).     

Expression of Aspergillus nidulans phy Gene in Nicotiana benthamiana Produces Active Phytase with Broad Specificities

A full-length phytase gene (phy) of Aspergillus nidulans was amplified from the cDNA library by polymerase chain reaction (PCR), and it was introduced into a bacterial expression vector, pET-28a. The recombinant protein (rPhy-E, 56 kDa) was overexpressed in the insoluble fraction of Escherichia coli culture, purified by Ni-NTA resin under denaturing conditions and injected into rats as an immunogen. To express A. nidulans phytase in a plant, the full-length of phy was cloned into a plant expression binary vector, pPZP212. The resultant construct was tested for its transient expression by Agrobacterium-infiltration into Nicotiana benthamiana leaves. Compared with a control, the agro-infiltrated leaf tissues showed the presence of phy mRNA and its high expression level in N. benthamiana. The recombinant phytase (rPhy-P, 62 kDa) was strongly reacted with the polyclonal antibody against the nonglycosylated rPhy-E. The rPhy-P showed glycosylation, two pH optima (pH 4.5 and pH 5.5), an optimum temperature at 45~55 °C, thermostability and broad substrate specificities. After deglycosylation by peptide-N-glycosidase F (PNGase-F), the rPhy-P significantly lost the phytase activity and retained 1/9 of the original activity after 10 min of incubation at 45 °C. Therefore, the deglycosylation caused a significant reduction in enzyme thermostability. In animal experiments, oral administration of the rPhy-P at 1500 U/kg body weight/day for seven days caused a significant reduction of phosphorus excretion by 16% in rat feces. Besides, the rPhy-P did not result in any toxicological changes and clinical signs.     

Biotechnological potential of industrial wastes for economical citric acid bioproduction by Aspergillus niger through submerged fermentation

Submerged citric acid (CA) bioproduction was carried out by Aspergillus niger NRRL‐567 using various industrial wastes, such as brewery spent liquid (BSL), lactoserum and starch industry water sludge. CA bioproduction was carried out by varying the temperature (25–35 °C), pH (3–5), addition of inducers, incubation time and supplementation with different proportions of apple pomace ultrafiltration sludge (APS). The results indicated that under the best conditions with 3% (v/v) methanol, the optimal concentration of 11.34 g L^?1 CA was recorded using BSL at pH 3.5 and temperature 30 °C after 120‐h incubation period. Supplementation of methanol resulted in an increase of 56% CA production. Meanwhile, under similar conditions, higher concentration of 18.34 g L^?1 CA was reported with the supplementation of BSL with 40% (v/v) APS having suspended solids concentration of 30 g L^?1. The present study demonstrated the potential of BSL supplemented with APS as an alternative cheap substrate for CA fermentation.