响应面法优化硇洲马尾藻多糖提取工艺

为优化硇洲马尾藻多糖提取工艺,在单因素试验基础上,选择提取温度,提取时间,料液比为自变量,多糖提取率为响应值,利用Box-Benhnken中心组合试验和响应面分析法,研究各自变量交互作用及其对硇洲马尾藻多糖得率的影响,模拟得到二次多项式回归方程的预测模型,并确定硇洲马尾藻多糖提取工艺最佳条件为:料液比为1:21(g/mL),提取温度为90.7℃,提取时间为5.14 h,在此条件下,硇洲马尾藻多糖得率达到6.39%。     

8株窖泥链霉菌对酸、乙醇的适应性研究

对分离自窖泥的8株放线菌进行鉴定,并考察其对窖泥环境中pH值、乙醇的耐受性.采用ISP2液体培养基在pH值2.0～3.5、乙醇浓度3%vol ～6%vol条件下培养链霉菌菌株,结果显示,分属于链霉菌属的8个种的8株放线菌均不能耐受pH值2.0及6%vol的乙醇浓度,均能耐受pH值3.5及3%vol的乙醇浓度,5株链霉菌在pH值3.0时可以生长,2株链霉菌可耐受5%vol乙醇,说明分离自窖泥的多数链霉菌对窖泥环境的适应能力较好,但不同种链霉菌的适应能力存在差异,窖泥环境在长期生产过程中对链霉菌属菌株具有一定的选择驯化作用.     

Production of lepidimoide by an endophytic fungus from polysaccharide extracted from Abelmoschus sp.: identification of the product and the organism producing it

A highly potent allelopathic factor, lepidimoide, was initially extracted from mucilage of germinated cress seeds. Polysaccharide extracted from okra (Abelmoschus esculentum Moench) is considered to have a similar structure to lepidimoide as its repeating unit. We therefore initiated the screening of enzymes capable of degrading okra polysaccharide into lepidimoide from endophytes. We discovered an endophytic fungal strain AHU9748 isolated from Coleus galeatus, which produced an oligosaccharide having similar properties to lepidimoide on thin layer chromatography. The physico-chemical data from ESI-MS, NMR spectra and other analyses also showed the purified product to be identical to lepidimoide. The strain AHU9748 was identified as a fungus belonging to the coelomycetes, closely related to the genus Colletotrichum, based on morphological characteristics and sequence analysis of the 18S rDNA and ITS region.     

Production and Stability Studies of a Neurotoxin Produced by Clostridium sp. RKD

ABSTRACT: A neurotoxigenic Clostridium sp. (RKD) isolated from intestine of decaying fish produced a neurotoxin that was neutralized by botulinum antitoxin (A+B+E) when tested by mouse protection bioassay. An amplicon of expected size (approximately 700 bp) was generated with primers specific for BoNT/B. Toxin was maximally released in the culture supernatant in the late stationary phase and was dependent on media composition. Growth was optimal in trypticase peptone yeast-extract glucose (TPYG) medium in a pH range of 7.5 to 8.0 and at a temperature between 35°C to 40°C while toxin production was optimum at 37°C (3 to 4 × 10³ minimum lethal dose per milliliter) without any protease treatment. There was no correlation between growth and toxin production when cells were grown in media containing different concentrations of NaCl (0% to 5%). Toxin in the culture supernatant was more stable (50% reduction at 50°C in 90 min) than the partially purified fraction. Toxicity was destroyed gradually after increasing the number of freeze-thaw cycles and was almost completely inactivated after 5 cycles. It was completely inactivated by overnight treatment of 1 N NaOH while it retained 1.5% activity with a similar treatment with 1 N HCl.     

短杆菌属产胆固醇氧化酶的发酵条件优化

短杆菌属(Brevibacterium sp.)是胆固醇氧化酶高产菌,对其进行底物诱导及发酵条件的优化,其最适培养基为蔗糖0.3%,酵母膏0.2%,蛋白胨0.3%,牛肉膏0.3%,K2HPO4 0.1%,MgSO4 0.05%,pH6.8。最适培养条件为接种量5%,24℃培养20h,通气量为50mL培养基/250mL三角瓶,200r/min。结果表明,胆固醇氧化酶的酶活力可达到2439U/L,比未优化前195U/L提高了12倍。在pH6.5和温度54℃条件下测得酶米氏常数Km值为7.1×10-5 mol/L。     

降烟碱节杆菌发酵条件研究

在12种发酵培养基、不同温度、pH和培养时间条件下,对细菌生长能力和降烟碱能力进行测定,初步筛选出适宜4#菌株生长的为11号培养基,在5号、7号、9号培养基中4#菌株降烟碱能力强;适宜9#菌株生长的为8号培养基,在7号、9号、11号、12号培养基中9#菌株的降烟碱能力较强。适宜4#和9#的培养条件分别为30℃、pH6.5、48h和35℃、pH6.5、56h。     

Application of brown-rot basidiomycete Fomitopsis sp. IMER2 for biological treatment of black liquor

Acid-producing fungi can be used for the treatment of black liquor by biological acidification for the precipitation of alkali lignin. The effects of alkali lignin obtained from black liquor on the growth, acid-producing capacity and glucose consumption of a brown-rot fungus Fomitopsis sp. IMER2 were investigated and the changes in the functional group in the resulting alkali lignin were analyzed by Fourier transform infrared (FTIR) spectroscopy. Although the presence of alkali lignin inhibited fungal growth and acid production, it enhanced glucose consumption. This suggests that the physiological response of this fungus to stress induced by alkali lignin is unique. Furthermore, the analysis of the effects of the resulting alkali lignin on the growth rate showed that several functional groups were evidently improved for the use of available resources.     

Characterisation of a novel laminarinase from Microbulbifer sp. ALW1 and the antioxidant activity of its hydrolysates

Laminarin and its derived oligosaccharides have diverse bioactivities. The β-1,3-glucanase in marine bacteria can be employed as a tool to digest laminarin in the cell wall of brown algae. Here, we cloned, expressed and characterised a β-1,3-glucanase (laminarinase), MaLamNA, from the previously characterised marine bacterium Microbulbifer sp. ALW1, phylogenetically distinct from the glycoside hydrolase families of characterised laminarinases. The recombinant laminarinase was heterologously expressed and purified from Pichia pastoris GS115 cells with a molecular mass of 57.3 kDa. MaLamNA exerted its hydrolytic activity specifically against laminarin, with the highest activity at 45 °C and pH 4.5–5.5, respectively, and demonstrated high stability against extreme acidic and alkaline pH exposure. The addition of reducing agent dithiothreitol could significantly enhance the activity of MaLamNA. The hydrolytic products of laminarin by MaLamNA exhibited more effective antioxidant activities than the undigested laminarin. These characteristics of MaLamNA provide clues to its industrial application for laminarin bioresource development.     

根霉ZM-10脂肪酶的分离纯化和活性部位的化学修饰

根霉ZM-10脂肪酶经过硫酸铵盐析、DEAE-Sepharose FF阴离子交换层析、SephardexG100凝胶过滤层析得到电泳纯脂肪酶,分子量约为42 ku,纯化倍数15.3倍,酶活回收率22.2%。采用NBS、EDC、DEPC、CH-T、PMSF、DTNB等6种化学修饰剂对根霉ZM-10脂肪酶进行化学修饰和底物保护实验,研究了其分子中氨基酸侧链基团与酶活性中心的关系。实验结果表明,酸性氨基酸(天冬氨酸?谷氨酸)残基、组氨酸残基、丝氨酸残基、色氨酸残基为根霉ZM-10脂肪酶活性的必需基团。酶分子中酸性氨基酸(天冬氨酸?谷氨酸)残基、组氨酸残基和丝氨酸残基位于根霉ZM-10脂肪酶的活性中心部位,而色氨酸残基在保持脂肪酶活性中起到重要作用,但其不位于根霉ZM-10脂肪酶的活性中心部位。     

Influence of indigenous yeasts on the fermentation and volatile profile of plum brandies

The aim of this study was to determine the influence of different yeasts isolated from fresh blue plum fruits (Aureobasidium sp.) and spontaneously fermenting plum musts (Kloeckera apiculata and Saccharomyces cerevisiae), as well as commercial wine and distillery strains, on the fermentation and chemical composition of plum brandies. Gas chromatography methods were used to detect major volatile components. The most rapid fermentation occurred in musts inoculated with S. cerevisiae. However, the highest concentration of ethanol was detected in samples after spontaneous fermentation (8.40% v/v). Plum brandies obtained after distillation contained from 66.3 (K. apiculata) up to 74.3% v/v ethanol (spontaneous fermentation). The samples after spontaneous fermentation were distinguished by a high content of acetoin, ethyl acetate and total esters, accompanied by a low level of methanol and fusel alcohols. Non-Saccharomyces yeasts were responsible for higher concentrations of esters and methanol, while S. cerevisiae strains resulted in increased levels of higher alcohols. It was also found that isolated indigenous strains of S. cerevisiae synthesized relatively low amounts of higher alcohols compared to commercial cultures. Samples obtained using the distillery strain of S. cerevisiae received the highest score (18.2) during sensory analysis and were characterized by a well-harmonised taste and aroma.