同步辐射软X射线Nk对产L-乳酸米根霉的诱变效应

采用同步辐射软X射线Nk对米根霉AS 3.3461菌株进行辐射诱变,研究其对米根霉致死、菌落形态、种子培养以及发酵培养等特性方面的影响。结果表明,致死率曲线呈现为典型的马鞍型曲线,辐射剂量为0.72 kGy时菌株致死率最低;软X射线Nk辐照对米根霉菌落形态有一定影响;种子培养特性和发酵培养特性的诱变前后对比表明,种子延滞期缩短了3h,突变株N-15的L-乳酸产量提高了25.6%,LDH活性提高了95.8%,ADH的活性降低了77.7%。     

米根霉发酵米糠工艺优化及其益生活性研究

为探究米根霉发酵米糠的最佳条件,本研究采用过热蒸汽对米糠作稳定化处理,以米根霉为菌种发酵稳定化米糠,以米糠可溶性膳食纤维(SDF)得率为指标,并对此条件下发酵米糠的体外益生活性进行评价。结果表明,过热蒸汽稳定米糠最佳条件为160℃、2 min,此条件下脂肪酶失活率为88.01%;甜香型米根霉发酵米糠的最佳条件为:以甜香型米根霉为发酵菌种,料液比1:2.25、接种量0.7%、发酵温度35℃、发酵时间24 h,此条件下SDF得率为5.83%,比未发酵米糠提高了近一半;与未发酵米糠相比,经甜香型米根霉发酵后的米糠,可有效促进双歧杆菌(对数值6.85)和乳酸杆菌(对数值5.23)的增殖;总短链脂肪酸含量提高37.30%,肠道益生活性显著增强(P<0.05)。本研究结果为改善米糠功能特性提供了依据,对今后米糠在发酵方面的应用具有一定的指导意义。     

米根霉丙烯醇抗性突变株的筛选及其发酵特性

本研究利用亚硝基胍(NTG)对米根霉As3.3462进行诱变,在含丙烯醇的YPD选择培养基上筛选获得11株ADH活力降低的突变株,其中mut-2突变株发酵液中乙醇含量比原始菌株降低35.2%,而乳酸含量提高了79.6%。对原始菌株和mut-2突变菌株在发酵过程中ADH与LDH活力变化分析表明,突变菌株的最大ADH活力比原始菌株降低了60.6%,而LDH活力与原始菌株相比略有提高。研究结果还表明,mut-2突变菌株与出发菌株相比,其生物量和对还原糖的利用速率均比原始菌株提高。     

抗稻瘟病突变品种的筛选及其利用研究

应用软 X 射线辐射诱变选育而成的高抗稻瘟病突变新品种申香粳4号,矮秆、抗病、优质。在所测的88个菌株中,能抗87个菌株,抗菌株率达98.86%。稻米品质优良,具爆玉米花香型香味。     

混菌发酵黑青稞甜醅工艺优化及品质特性研究

为改善单一菌株发酵制备青稞甜醅的风味与口感,提高其质量品质,本试验采用米根霉和酵母菌为发酵菌株,以氨基酸态氮含量及感官评分为指标,确定混菌发酵黑青稞甜醅的最佳工艺条件,并比较单一菌株发酵黑青稞制品与混菌发酵黑青稞制品中酚类物质含量、抗氧化活性及风味物质组成的差异。结果表明,混菌发酵黑青稞制品最佳发酵条件为:发酵温度33℃,发酵时间48 h,菌种比例(酵母菌J7∶米根霉)1∶1.20,接种量6.81%,在此条件下混菌发酵黑青稞制品的氨基酸态氮含量为9.32 mg·100 g^-1,感官评分为95.48分。与单一菌株发酵黑青稞制品相比,混菌发酵黑青稞的黄酮含量(32.22 mg·100 g^-1)、 多酚含量(230.68 mg·100 g^-1)及DPPH自由基清除能力(95.03 μmol·L^-1)显著提高。气相色谱-质谱(GC-MS)分析结果表明,酵母菌单独发酵的黑青稞中共检出33种挥发性风味物质,米根霉单独发酵的黑青稞中共检出41种挥发性风味物质,混菌发酵的黑青稞中共检出46种挥发性风味物质,其中酯类和醇类是3种发酵方式黑青稞制品的主要风味组分。混菌发酵黑青稞的醇类、酯类和酸类种类及含量均显著高于其余两种发酵方式,其相对含量分别达到59.09%、29.44%和6.46%,风味更丰富。综上分析,混菌发酵使黑青稞制品在功能及风味方面具有一定的优势。本研究结果为混菌发酵黑青稞制品的开发和应用奠定了基础。     

高压电晕电场对黄霉素产生菌诱变效应

利用高压电晕电场诱变技术对黄霉素产生菌08-28进行微生物代谢产量和活性的研究。结果表明:链霉素对黄霉素产生菌的最低抑制浓度为0.75U/ml;随着电场处理剂量的增大菌种致死率也随之增大,诱变电压为21kV,针尖距离为8mm,诱变时间为4min时是最佳诱变条件,此时致死率在68%,菌落形态呈放射状褶皱。通过筛选,获得8株突变高产菌株,其中产黄霉素能力最高的是原始菌株的1.92倍。     

抗马铃薯晚疫病菌的粘细菌菌株X6-Ⅱ-1的分离鉴定、拮抗活性及发酵条件的优化

马铃薯晚疫病(potato late bright)是马铃薯(Solanum tuberosum)最为严重的病害,其病原菌为致病疫霉(Phytophthora infestans)。为了从土壤样品中筛选出对马铃薯晚疫病具有拮抗作用的粘细菌,本实验利用大肠杆菌(Escherichia coli)划线诱导法从土样中分离菌株,通过平板对峙法筛选具有抗马铃薯晚疫病菌活性的粘细菌,结合形态观察、生理生化特征和16S r RNA基因序列分析确定菌株的分类地位。采用称重法测定菌株生长曲线,滤纸片法确定活性物质的分布,并通过单因素分析与正交优化相结合的方法对菌株的发酵参数进行了研究。共从土壤样品中共分离得到5株粘细菌,其中有3株对致病疫霉表现出拮抗活性,菌株X6-Ⅱ-1的拮抗活性最强,致病疫霉菌丝生长边缘与菌株X6-Ⅱ-1菌饼最短距离可达到5 mm,经鉴定为叶柄粘球菌(Myxococcus stipitatus)。该菌株可以杀死并溶解大肠杆菌,且可以抑制枯草芽胞杆菌(Bacillus subtilis)的生长。拮抗致病疫霉的活性物质主要存在于细胞外,产活性物质的最适发酵条件为:接种量10%,摇床转速180 r/min,培养温度为30℃,发酵时间为7 d。粘细菌菌株X6-Ⅱ-1能够产生抗马铃薯晚疫病菌的活性物质,具有开发成抗马铃薯晚疫病生物农药的潜在价值。本研究为活性物质的分离鉴定以及抗马铃薯晚疫病农药的研发提供基础数据。     

辐照木霉菌株的生物学效应研究

本文研究了6 0 Coγ射线辐照木霉 (Trichodermaspp)菌株的诱变效果。结果表明 :在γ射线的作用下 ,木霉分生孢子萌发率显著降低 ,孢子萌发时间加长 ,部分萌发孢子菌丝生长畸形 ;随着照射剂量的增加 ,木霉孢子存活率急剧下降 ,剂量效应曲线符合一次击中模型S =e-λD;诱变效应方程为 :Y =-0 940e1 2 9x,r=0 953 3 (r0 0 1=0 765)。依各菌株对生长速度、产孢量、开始产孢时间、菌落颜色等变异情况 ,初筛得到变异菌株 1 3 0株。     

辐照对糖多孢红霉菌M的诱变效应

采用不同剂量的60Co γ射线辐照糖多孢红霉菌M孢子,并采用不同的菌种保护剂,以研究60Coγ射线对糖多孢红霉菌M的诱变效应。结果表明,使用20%甘油吐温保护效果较好,可作为诱变保护剂;当辐照剂量为400 Gy时,致死率为90%左右可作为适宜诱变剂量。诱变处理后利用链霉素抗性突变选育得1203株链霉素抗性突变株。其中5株形态发生较明显的变化,形态变异率为4.16‰;2株突变株的发酵液具有抑菌活性,正突变率为1.67‰。     

137 Cs-γ辐射对葡萄种子发芽和幼苗生长及叶绿素荧光特性的影响

为了研究核辐射技术在诱变葡萄种子上的应用,本试验用不同剂量的137Cs-γ射线辐射处理醉金香葡萄成熟干种子,并将其培养成实生苗,研究了不同辐射剂量对葡萄种子发芽、幼苗生长、幼株的叶绿素荧光参数等生物学效应的影响,从而确定葡萄成熟种子的适宜辐照剂量,以期为开展葡萄诱变育种工作奠定理论基础.结果表明137Cs-γ射线辐射使种子发芽率、发芽势和发芽指数下降,平均发芽天数增加,出苗率、成苗率下降,幼苗株高变矮,茎杆变细、木质化时间延迟,黄化苗增多,且随辐射剂量增加影响加大.1377Cs-γ射线对葡萄叶绿素荧光参数的影响比对种子的小,对各荧光参数的影响也不同,且与辐射剂量不呈线性关系,高剂量(50Gy)的处理各荧光参数与对照差异显著,影响较大.综合上述各项指标,认为20～40Gy为醉金香葡萄γ射线辐射诱变较为合适的剂量范围.     

Endophytic fungi associated with Mediterranean plants as a source of mycelium-bound lipases

A screening of endophytic fungi isolated from Mediterranean plants rendered a mycelium-bound lipase from a strain of Rhizopus oryzae that catalyzed the esterification of fatty acids in isooctane. The influence of various factors (water content, temperature, and pH) on ester synthesis was investigated. Catalytic activity was inversely correlated with water content. This enzyme was active over the entire pH range studied, from pH 3 to pH 8, and activity was maximal at pH 4 and pH 7. The enzyme was thermostable, with maximal activity at 60 degrees C.     

Fermentation as a bio-process to obtain functional soybean flours

The effect of fermentation on the antioxidant compounds [vitamins C and E, total phenolic compounds (TPC), and reduced glutathione (GSH)], and antioxidant capacity [superoxide anion scavenging activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), inhibition of phosphatidylcholine (PC) peroxidation, and Trolox equivalent antioxidant capacity (TEAC)] of soybean (Glycine max cv. Merit) was studied. Fermentation was carried out in solid state in cracked seeds inoculated with Aspergillus oryzae, Rhizopus oryzae, Bacillus subtilis, and Lactobacillus plantarum and in liquid state either in cracked seeds or milled soybean flours fermented naturally by only the microorganisms present in the seeds or by inoculation with L. plantarum. Vitamin C was not detected in the studied samples. Fermentation caused a decrease in vitamin E activity, except when cracked seed was fermented with A. oryzae, R. oryzae, or B. subtilis that increased 31, 30, and 89%, respectively. Fermentation produced an increase in TPC content and did not affect or reduce the GSH content. Fermentation decreased SOD-like activity drastically, while PRTC increased except when it was carried out naturally in cracked seed. TEAC values rose sharply when soybeans were fermented with B. subtilis. Processed soybean extracts inhibited PC peroxidation in comparison with the control assay. On the basis of the results obtained, the relative contributions of vitamin E, TPC, and GSH to antioxidant capacity were calculated and results showed a very high TPC contribution and a low contribution of GSH and vitamin E activity. Optimum results for functional soybean flours were achieved when fermentation was carried out with B. subtilis inoculum.     

不同粮仓中稻谷优势霉菌的鉴定及差异性分析

为了研究稻谷中优势霉菌的种类、数量以及变化趋势,本试验以湖南省3个不同地区稻谷为研究对象,通过初步判断各粮仓优势霉菌,结合分子生物学方法对其ITS 序列进行分析,并对稻谷霉菌进行带菌量测定。结果表明,3个粮仓中,金牛仓稻谷中优势菌株分别为根霉、米曲霉、毛霉、黑曲霉、烟曲霉、黄曲霉、白曲霉、桔青霉;金山仓稻谷中优势菌株分别为根霉、米曲霉、毛霉、黑曲霉、黄曲霉、白曲霉、桔青霉;银光仓稻谷中优势菌株分别为根霉、米曲霉、毛霉、黑曲霉、黄曲霉。3个粮仓中根霉数量分布均表现为上层>下层>中层,米曲霉为中层>上层>下层,毛霉、黑曲霉、黄曲霉的分布为上层>中层>下层,同一霉菌在同一粮仓不同粮层间数量存在显著差异(P<0.05);随着储藏时间的延长,米曲霉、黑曲霉、黄曲霉数量逐渐减少,根霉、毛霉数量增加,同一霉菌在不同储藏时间霉菌数量存在差异显著(P<0.05)。本研究结果为稻谷安全储藏与霉菌防控提供了一定的理论依据。     

Enzymatic formation of ether linkage producing shoyuflavones from genistein and (+/-)-trans-epoxysuccinic acid

The production mechanism of shoyuflavones, conjugated ethers of isoflavones with tartaric acid and isolated from fermented soy sauce, was studied. In the high molecular weight fraction of the culture extract of Aspergillus oryzae, genistein was transformed into shoyuflavone B in the presence of (+/-)-trans-epoxysuccinic acid but not in the low molecular one. Asp. sojae and Asp. tamarii showed high activity similar to Asp. oryzae but none of Asp. niger, Rhizopus oligosporus, and Mucor praini did. The contents of epoxysuccinic acids in the starting materials of soy sauce and the cultures of various Asp. fungi were determined as dimethyl 2-chloro-3-hydroxysuccinate derivatives by GC-MS. Although epoxysuccinic acids were contained in Asp. oryzae, Asp. sojae, and Asp. tamarii cultures, they were not found in soybeans and wheat. A possible producing mechanism for shoyuflavones by enzymatically conjugating isoflavones to (+/-)-trans-epoxysuccinic acid with ether linkage was suggested.     

3株抗水稻和荔枝病原菌的海洋真菌的分离鉴定

本研究采用平板对峙法和菌丝块法测定分离自广西北部湾的3株海洋真菌MF-3、MF-6和MF-13的抗荔枝和水稻病原菌抗菌活性,并通过形态学观察结合分子生物学方法鉴定其分类地位。结果表明,当采用平板对峙法时,3株海洋真菌对荔枝霜疫霉、炭疽病菌、水稻纹枯病菌和水稻稻瘟病菌等4种指示菌的菌丝生长均有强的抑制作用;菌丝块法试验结果发现3株真菌的发酵液均对荔枝霜疫霉菌有很好的抑制效果,菌丝相对生长抑制率分别达到94.29%、98.16%和86.37%,对水稻纹枯病病原菌抑制效果次之,其菌丝相对生长抑制率达到80%左右;其中,MF-3菌株对荔枝霜疫霉、炭疽菌、水稻纹枯病菌和水稻稻瘟病菌都有较好的抑制作用,MF-6对霜疫霉、水稻纹枯病菌和水稻稻瘟病菌也有较好的抑菌效果和广谱性。结合形态学观察和ITS rDNA序列比对分析,初步将MF-3鉴定为布雷正青霉（Eupenicillium brefeldianum）,MF-6为微紫青霉（Penicillium janthinellum）,MF-13为短棒曲霉（Aspergillus clavatonanicus）。耐盐度试验结果表明这3株真菌是适应海洋环境的兼性海洋真菌。该3株海洋真菌表现的抗真菌活性,对应用于荔枝或水稻病害的生物防治和新型农药的研发有潜在的应用价值。     

Immunoreactivity and amino acid content of fermented soybean products

Food allergy has become a public health problem that continues to challenge both the public and the food industry. The objective of this research was the detection and quantification of the major human allergenic soy proteins and to study the reduction in immunoreactivity and improvement of amino acid content after fermentation of soybean flour. Fermentation was carried out in the solid state of cracked seeds inoculated with Aspergillus oryzae, Rhizopus oryzae, and Bacillus subtilis and in the liquid state of milled soybean flours fermented naturally by microorganisms present only in the seeds or by inoculation with Lactobacillus plantarum. ELISA and Western blot were used to quantify IgE antibody response, and HPLC was used to identify and quantify total amino acids. L. plantarum fermented soy flour showed the highest reduction in IgE immunoreactivity (96-99%) depending upon the sensitivity of the plasma used. Among the solid fermented products, the lowest reduction in immunoreactivity was obtained when mold strains, R. oryzae and A. oryzae, were used (66 and 68%, respectively, for human plasma 97.5 kUA/L). Among the solid fermented products, those inoculated with B. subtilis yielded a 81 and 86% reduction in immunoreactivity against both human plasma 97.5 IgE kUA/L and human pooled plasma samples, respectively. When soybean was subjected to liquid fermentation, most of the total amino acids increased significantly ( p < or = 0.05). In solid fermentation with R. oryzae, only Ala and Thr content improved. Fermentation can decrease soy immunoreactivity, and there is potential of developing nutritious hypoallergenic soy products.     

Development of an oat-based biorefinery for the production of L(+)-lactic acid by Rhizopus oryzae and various value-added coproducts

A novel oat-based biorefinery producing L(+)-lactic acid and various value-added coproducts (e.g., beta-glucan, anti-irritant solution) is proposed. Pearling is employed for sequential separation of bran-rich fractions for the extraction of value-added coproducts. Lactic acid production is achieved via fungal fermentation of Rhizopus oryzae on pearled oat flour. Maximum lactic acid concentration (51.7 g/L) and starch conversion yield (0.68 g/g) were achieved when an oat flour concentration of 116.5 g/L was used. Oxygen transfer played a significant role with respect to lactic acid production and starch conversion yield. Rhizopus oryzae produced a range of enzymes (glucoamylase, protease, phosphatase) for the hydrolysis of cereal flour macromolecules. Enzyme production during fungal fermentation has been reported. The proposed biorefining strategy could lead to significant operating cost reduction as compared to current industrial practices for lactic acid production from pure glucose achieved by bacterial fermentations.