黑曲霉HU53菌株产酸性蛋白酶的条件和酶学性质

本文报道了黑曲霉（Aspergillus niger)HU53菌株产酸性蛋白酶的固体发酵条件优化和酶学特性的研究结果。适合该菌株的最佳固体发酵培养基为麦麸38．8％，豆粕9．7％，NH4NO31．5％和水50．0％，最佳起始pH5．5。在28℃下发酵50h后酸性蛋白酶的比活力达到93．8U／g，且在52h发酵期内基本不产孢子。该酸性蛋白酶的最佳反应酸碱度为pH3．0，最佳反应温度为50℃，对酪蛋白的Km为2．8016mg／ml，在40℃下保温180min后相对酶活力为93．50％。2．0mmol／L的Cu^2 和Mn^2 对该酸性蛋白酶具有极其显著的激活作用，相对酶活力分别为对照的171．9％和121．1％；而相同浓度的Fe^3 和Ca^2 则对其表现出强烈的抑制作用，相对酶活力分别仅为对照的62．5％和44．6％。     

Induction of chitinase in response to Aspergillus infection in sorghum (Sorghum bicolor (L) Moench)

Chitinase is an antifungal protein which is induced in higher plants during infection and stress. In this paper the induction of chitinase activity in response to infection by Aspergillus parasiticus (NRRL 2999) in six sorghum (Sorghum bicolor (L) Moench) genotypes and its association with aflatoxin production were investigated. Chitinase was induced in all six genotypes (two each of red, yellow and white sorghum) when infected by A parasiticus (NRRL 2999). The induction of chitinase activity was highest in the white cultivars, followed by the yellow and red genotypes, compared with healthy grains. In the white cultivars the chitinase activity increased on the 6th and 9th days after infection and was four‐ to fivefold higher than in healthy grains. The total aflatoxin produced was lower in the red genotypes than in the yellow and white genotypes. The white genotypes showed maximum total aflatoxin production at 6 days after infection. The aflatoxins produced in the white genotypes were comparable to those in the red genotypes. There was a significant positive correlation between chitinase activity and aflatoxin production in red sorghum (r² = 0.600, p ≤ 0.001) and white sorghum (r² = 0.411, p ≤ 0.001). Maximum chitinase activity was observed on day 12 in all genotypes under healthy conditions. Copyright © 2004 Society of Chemical Industry     

Inhibitory effects of anethole and eugenol on the growth and toxin production of Aspergillus parasiticus

The antifungal and antiaflatoxigenic activity of anethole and eugenol which are active components of commonly used spices was studied against two strains of Aspergillus parasiticus. Anethole, up to concentration of 400 μg/ml where complete inhibition was observed, delayed growth and reduced mycelial weight but it showed a stimulative effect on the toxin production of both strains. At a concentration of 300 μg/ml, eugenol inhibited the growth of both strains; levels of eugenol below 200 μg/ml enhanced production of aflatoxin particularly by A. parasiticus NRRL 299.     

Identification of Aspergillus tubingensis strains responsible for OTA contamination in grapes and wine based on the acyl transferase domain of a polyketide synthase gene

Restriction digestion analysis of the acyl transferase (AT) domain sequences of a polyketide synthase (PKS) gene was tested as a rapid method to identify isolates of Aspergillus tubingensis from grapes. Restriction endonuclease digestion of PKS products using the endonucleases Bcc I, Hae III, Hpa II, Mbo I and Taq I distinguished five types of restriction fragment length polymorphism (RFLP). Ochratoxigenic isolates were only identified within RFLP-types I and III. The RFLP assay is proposed as a rapid and easy method to identify A. tubingensis isolates from grapes. Amino acid sequences of AT domains from representative A. tubingensis isolates of the RFLP types obtained were aligned and analysed using phylogenetic methods. A comparison was also made with reference strains of Aspergillus section Nigri . Most of the A. tubingensis strains clustered into two distinct groups Gr1 and Gr2 with the exception of two isolates that remained unclustered. These results support the intraspeficific variability within A. tubingensis species reported using other techniques.     

Solid-state fermentation of Lathyrus sativus seeds using Aspergillus oryzae and Rhizopus oligosporus sp T-3 to eliminate the neurotoxin β-ODAP without loss of nutritional value

The presence of a neurotoxic amino acid, 3-N-oxalyl-L-2,3-diaminopropanoic acid (β-ODAP), in the seeds of grass pea (Lathyrus sativus) is the cause of an irreversible spastic paraparesis, neurolathyrism, after over-consumption. This is a negative factor in an otherwise very tasty, nutritious, easily cultivated and hardy food crop in Asia and Africa. The reduction of β-ODAP to less than 10% of the original content has been reached by fermenting L sativas Jamalpur seeds with Aspergillus oryzae NRRL 1988 for 48 h, followed by fermentation with Rhizopus oligosporus sp T-3 for 48 h. Other nutritional qualities were also improved in the fermented seed meal: increased content of protein, higher amino acid scores for sulphur-containing and aromatic amino acids, better resistance to high temperature and to oxidation, and drastic reduction of the flatulence factors.     

Effect of pH on ochratoxin A production by Aspergillus niger aggregate species

The effect of pH (2-10) on growth and ochratoxin A (OTA) production by 12 Aspergillus niger aggregate strains was studied in two culture media: Czapek yeast autolysate agar (CYA) and yeast extract sucrose agar (YES), over 30 days. The strains were selected to include different sources, different reported abilities to produce OTA and different ITS-5.8S rDNA RFLP patterns. YES was a better culture medium than CYA for OTA production. In this medium, OTA was produced from pH 2 or 3 to 10 depending on the strain. The results show the ability of A. niger aggregate strains not only to grow, but also to produce OTA over a wide pH range. The results will lead to a better understanding of the role of A. niger aggregate strains in the OTA contamination of several food commodities.     

贵州茯茶散茶发花条件优化及微生物多样性分析

贵州是我国茶树品种资源最丰富的省份之一,茶园面积大,但经济产值低,夏秋茶浪费严重,品种结构单一。该研究以贵州省安顺市黑毛茶为供试原材料,人工接种茯砖茶“金花”发酵剂,优化散茯茶发花工艺条件,并开展了理化指标变化及真菌多样性研究。通过单因素试验及正交优化试验分析不同因素对黑毛茶散茶发花的影响。结果表明,其最佳发花条件为：含水量30%、温度24 ℃、发酵时间9 d及接种量1:100（m/m）。在此发花条件下,成品散茯茶冠突曲霉孢子数为3×108 CFU/mL,水分、总灰分、茶多酚、游离氨基酸、水浸出物含量分别为3.0%、6.8%、12.0%、2.4%、42.6%。经过上述优化后,茶汤色明亮,金花茂盛,香气纯正,滋味醇和。通过高通量测序技术分析不同发酵时间下散茯茶的真菌多样性。结果表明,发酵3 d后,曲霉属（Aspergillus）相对丰度达到98.8%。该研究优化了贵州散茶发花的工艺条件,对提高茯砖茶品质具有重要意义,为其他茶叶散茶发花工艺提供参考。     

高糖化力菌种的筛选及诱变育种

固体曲中精化酶能将淀粉水解为葡萄精,进而被微生物发酵生成酒精.以分离筛选出的1株产糖化酶的黑曲霉(酶活265.21U/g干曲)为出发菌,经紫外线及原生质体紫外线复合诱变育种,获得1株糖化酶活力较高、生产性能稳定的菌株,其酶活达到3367.79U/g干曲,比原始菌株提高1169.86%.     

黑曲霉原生质体诱变及其产纤维素酶条件研究

采用紫外、硫酸二乙酯及复合诱变3种方法对黑曲霉原生质体进行诱变育种,以提高其酶活。结果表明,采用复合诱变效果最好,复合诱变所得菌株的酶活比出发菌株大幅度提高;应用单因素实验优化突变菌株的产酶条件,其最佳产酶条件培养基为:5%蔗糖、1.4%蛋白胨、0.2%磷酸二氢钾、0.05%硫酸镁,在250 mL摇瓶中装液60 mL,10%接种量、29℃下200 r/min培养96 h,酶活达到最大。