The AFLATOX® Project: Approaching the Development of New Generation,Natural-Based Compounds for the Containment of the Mycotoxigenic Phytopathogen Aspergillus flavus and Aflatoxin Contamination

The control of the fungal contamination on crops is considered a priority by the sanitary authorities of an increasing number of countries, and this is also due to the fact that the geographic areas interested in mycotoxin outbreaks are widening. Among the different pre- and post-harvest strategies that may be applied to prevent fungal and/or aflatoxin contamination, fungicides still play a prominent role; however, despite of countless efforts, to date the problem of food and feed contamination remains unsolved, since the essential factors that affect aflatoxins production are various and hardly to handle as a whole. In this scenario, the exploitation of bioactive natural sources to obtain new agents presenting novel mechanisms of action may represent a successful strategy to minimize, at the same time, aflatoxin contamination and the use of toxic pesticides. The Aflatox^® Project was aimed at the development of new-generation inhibitors of aflatoxigenic Aspergillus spp. proliferation and toxin production, through the modification of naturally occurring molecules: a panel of 177 compounds, belonging to the thiosemicarbazones class, have been synthesized and screened for their antifungal and anti-aflatoxigenic potential. The most effective compounds, selected as the best candidates as aflatoxin containment agents, were also evaluated in terms of cytotoxicity, genotoxicity and epi-genotoxicity to exclude potential harmful effect on the human health, the plants on which fungi grow and the whole ecosystem.     

常压室温等离子体诱变选育高产曲酸米曲霉

以米曲霉KA2308作为出发菌株,首次利用常压室温等离子体诱变技术选育高产曲酸菌株,通过96孔板高通量筛选获得一株曲酸生产菌KA1515,曲酸产量提高到47.28g·L-1,比出发菌株提高了31.48%,经过多次传代,曲酸产量仍比较稳定。表明采用常压室温等离子体诱变能有效提高米曲霉曲酸产量。     

微生物絮凝剂的絮凝特性研究

从活性污泥中分离出1株产高效絮凝剂微生物AspergillusM-25,利用其发酵液进行絮凝特性实验。实验结果表明:(1)该絮凝剂对250μm,4g/L,pH为7的高岭土悬浊液有良好的絮凝效果,絮凝率>97%;(2)菌株M-25产生的活性成分95%存在于上清液中,只有约5%存在于洗涤后的菌细胞中,未接种的培养液中不含絮凝成分;⑶影响生物絮凝剂絮凝效果的因素主要有絮凝剂的投加量,高岭土悬浊液的pH、浊度、粒度以及金属离子的添加;⑷利用该生物絮凝剂处理活性炭粉末溶液和土壤悬浊液,也取得较好的处理效果,显示了絮凝范围的广泛性。     

高碱性铜尾矿的黑曲霉生物浸出

以湖南某高碱性铜尾矿为研究对象,进行了黑曲霉摇瓶浸出实验,研究了浸出时间和PSA培养基成分、含量对铜浸出的影响. 结果表明,浸出时间为7 d时,铜浸出率最高,随浸出时间延长,铜浸出率显著下降. PSA培养基中的马铃薯和蔗糖含量对铜浸出有明显影响,马铃薯含量为200 g/L、蔗糖含量为20 g/L时,铜浸出率最高. 在接种量0.02%(j)、矿浆浓度50 g/L、温度30℃、转速200 r/min、浸出时间7 d、马铃薯和蔗糖含量分别为200 g/L和20 g/L条件下,铜浸出率可达79.03%.     

酱油曲菌酶系及其产酶能力

研究了Ａｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅ Ａｓ．３９５１,ＪＰ－ＤＱ－１和ＪＰ－Ｗ的酶系和在不同培养基上的产酶能力。结果表明：上述三种菌在小麦曲、麦麸曲、米曲、浓酱油曲和白酱油曲上均可产生α－淀粉酶、糖化酶、纤维素外切酶、果胶酶和蛋白酶。ＪＰ－Ｗ在小麦曲、麦麸曲和白酱油曲上产酶能力最佳;ＪＰ－ＤＱ－１在浓酱油曲上产酶能力较强;Ａｓ．３９５１在米贡上产酶能力强。建议用ＪＰ－ＤＱ－１生产浓酱油,用Ｐ     

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。     

海洋来源真菌烟曲霉(Aspergillus fumigatus)次级代谢产物研究(Ⅰ):生物碱

对一株海洋来源的微生物中的次级代谢产物进行了研究,从中分离并鉴别了5个喹唑啉生物碱类化合物(1~5),分别为fumiquinazoline G(1),fumiquinazoline F(2),fumiquinazoline A(3),fumiquinazoline C(4)和fumiquinazoline D(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.     

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     

菊粉酶产酶菌株筛选与鉴定

从徐州沛县河口镇秦庄村牛蒡种植基地取得的土壤样本中,筛选出产菊粉酶的菌株,对从土壤中分离到的40株产菊粉酶的各类菌株进行酶活的测定．通过透明圈法初筛及摇瓶复筛,获得产菊粉酶能力较高的霉菌3株,为C122803、D081506和D081513．这三株菌株的酶活分别达到1．411u／mL、1．895u／mL、1．792u／mL,且其I／S值各达到1．1821、1．6806、1．3483．其中D081506的I／S值最大．对这三株菌株进行初步的微生物分类鉴定,符合黑曲霉的形态特征,初步鉴定为黑曲霉．