Purification and characterization of two low molecular weight endoglucanases produced by Penicillium occitanis mutant pol 6

Two endoglucanases (EGs), EG A and EG B, were purified to homogeneity from Penicillium occitanis mutant Pol 6 culture medium. The molecular weights of EG A and EG B were 31,000 and 28,000 kDa, respectively. The pI was about 3 for EG A and 7.5 for EG B. Optimal activity was obtained at pH 3.5 for both endoglucanases. Optimal temperature for enzyme activity was 60 degrees C for EG A and 50 degrees C for EG B. EG A was thermostable at 60 degrees C and remained active after 1 h at 70 degrees C. EGs hydrolyzed carboxymethylcellulose, phosphoric acid swollen cellulose, and beta-glucan efficiently, whereas microcrystalline cellulose (Avicel) and laminarin were poorly hydrolyzed. Only EG B showed xylanase activity. Furthermore, these EGs were insensitive to the action of glucose and cellobiose but were inhibited by the divalent cations Hg2+, Co2+, and Mn2+.     

Control Efficacy of Salicylic Acid Microcapsules against Postharvest Blue Mold in Apple Fruit

Salicylic acid (SA) is a natural inducer of disease resistance in fruit, but its application in the food industry is limited due to low water solubility. Here, SA was encapsulated in β-cyclodextrin (β-CD) via the host–guest inclusion complexation method, and the efficacy of SA microcapsules (SAM) against blue mold caused by Penicillium expansum in postharvest apple fruit was elucidated. It was observed that SAM was the most effective in inhibiting the mycelial growth of P. expansum in vitro. SAM was also superior to SA for control of blue mold under in vivo conditions. Enzyme activity analysis revealed that both SA and SAM enhanced the activities of superoxide dismutase (SOD) and phenylalanine ammonia lyase (PAL) in apple fruit, whereas SAM led to higher SOD activities than SA. Total phenolic contents in the SAM group were higher than those in the SA group at the early stage of storage. SAM also improved fruit quality by retarding firmness loss and maintaining higher total soluble solids (TSS) contents. These findings indicate that microcapsules can serve as a promising formulation to load SA for increasing P. expansum inhibition activity and improving quality attributes in apple fruit.     

A New Cytotoxic Compound from Penicillium auratiogdseum, Symbiotic or Epiphytic Fungus of Sponge Mycale plumose

A new compound, （S）-2, 4-dihydroxy-1-butyl （4-hydroxy） benzoate （1）, and a known compound, fructigenines A （2）, were isolated from fungus Penicillium auratiogriseum derived from sponge Mycale plumose, by bioassay-guided fractionation. Their structures were established by spectroscopic and chemical methods. Both compounds show.ed cytotoxic activity against tsFT210 cells.     

解磷青霉菌的筛选及其对农作物和牧草的增产作用

青霉菌是大多数土壤微生物群体中的成员。许多青霉菌具有生物防治功能、分解土壤矿物功能(包括解磷作用)以及能够分泌植物生长刺激物质。研究利用这些真菌来增加作物产量由此受到注意。本文介绍从土壤中分离、鉴定青霉菌、青霉菌的解磷作用以及对植物生长的刺激作用的研究方法以及策略。从澳大利亚的14种不同的农业土壤中分离并鉴定了解磷的青霉菌。某些菌株具有商业化价值,特别适合在碱性土壤中使用。这些菌株包括一个Penicillium bilaiae以及尚未鉴定的青霉菌种类。     

真菌吸附重金属离子的研究

利用黑曲霉和简青霉制备生物吸附剂,研究了它们对重金属Pb2 离子和Cd2 离子的吸附、解吸行为以及实验条件对吸附的影响,包括吸附剂用量、溶液pH值、吸附时间以及共存离子等因素.结果表明,黑曲霉和简青霉吸附Pb2 离子的最适pH值均为5,吸附Cd2 离子时均为3.二者对Pb2 离子的吸附均在4 h达到平衡,吸附量分别为29.07 mg·g-1和36.65 mg·g-1.Cd2 离子吸附也在约4 h达到最大吸附量,分别为26 mg·g-1和26.5mg·g-1.溶液中Zn2 离子和Cd2 离子的存在都会降低Pb2 离子的吸附量.2种吸附剂对Pb2 离子的吸附都符合Langmuir等温线模型,而对Cd2 离子的吸附都较为符合Freundlich等温线模型.1 mol/L HNO3对吸附有Pb2 离子的黑曲霉和简青霉进行解吸,解吸率分别可达77.4%和92.3%.     

重组草酸青霉生淀粉糖化酶的酶学特性鉴定

【目的】了解在毕赤酵母中表达的来源于草酸青霉(Penicillium oxalicum)GXU20的重组生淀粉糖化酶的酶学特性。【方法】用3,5-二硝基水杨酸法测定pH值、温度、金属离子和化学试剂对纯化的重组生淀粉糖化酶的活力的影响,同时测定酶的底物特异性、酶对生淀粉的吸附能力以及酶对不同生淀粉的水解效率等,用高效液相色谱法对该酶水解生木薯淀粉的产物进行分析鉴定,用扫描电子显微镜观察重组酶对不同生淀粉颗粒的水解方式。【结果】重组生淀粉糖化酶rPoGA15A的最适pH值为4.5,最适温度为65℃,在pH值为2.0~10.0时,重组酶具有较好的pH耐受性,温度小于50℃时,酶的稳定性好。除Ag~+、Cu~(2+)和SDS之外,其他大部分金属离子和化学试剂对重组酶的酶活力影响不大。重组生淀粉糖化酶对大米和玉米生淀粉的活性较高,对木薯和马铃薯生淀粉的活性次之,重组生淀粉糖化酶rPoGA15A对不同生淀粉的吸附能力与其对相应不同生淀粉的水解活性大小呈正相关。扫描电子显微镜观察表明重组生淀粉糖化酶对不同生淀粉颗粒的降解作用明显,水解方式各有特点。高效液相色谱分析表明该重组生淀粉糖化酶水解生木薯淀粉的产物仅有葡萄糖。重组生淀粉糖化酶rPoGA15A在40℃下对大米和玉米生淀粉水解72h后水解率分别达到86.5%和71.9%。【结论】重组生淀粉糖化酶具有广泛的pH耐受性,对生淀粉具有高水解活性,在生淀粉的水解和生料同步糖化发酵生产酒精中有一定的应用潜力。     

酸性木聚糖酶交联酶聚集体的制备条件优化

交联酶聚集体法是一种新型的无载体酶固定化方法,为提高酸性木聚糖酶的稳定性,使用该法固定化微紫青霉产酸性木聚糖酶,制备无载体固定化木聚糖酶,并对其制备条件进行优化。结果表明,优选的制备条件为将质量浓度0.36mg/mL的酸性木聚糖酶粗酶液在冰水浴中经饱和质量分数为85%的硫酸铵沉淀30min后,于40℃,加入终体积分数为0.14%的戊二醛,交联4h可获得较高活性的交联酶聚集体,酶活保留率达42.2%。这有助于酸性木聚糖酶更好地在工业中应用。     

Mode of action and synergism of cellulases fromPenicillium funiculosum

A 1,4-β-d-glucan cellobiohydrolase (EC 3.2.1.91) and l,4-β-d-glucan glucanohydrolase (EC 3.2.1.4) were purified from the culture filtrates ofPenicillium funiculosum by using preparative isoelectric focusing. Both the enzymes were homogeneous on polyacrylamide gel with and without sodium dodecyl sulphate. The mol wt of the cellobiohydrolase and endoglucanase were 14,400 and 25,000 respectively. The purified enzymes were free of β-glucosidase activity. Acting in isolation, the cellobiohydrolase had little capacity for solubilizing Avicel or Walseth cellulose, but showed increased rates of hydrolysis when combined with endoglucanase. Cellobiose inhibition (50%) was observed in the initial rate of the hydrolysis of Walseth cellulose. It was also observed that cellobiohydrolase initiates the attack on crystalline cellulose. † NCL communication no. 3898.     

P(3-co-4)HB解聚酶的分离纯化及酶学性质研究

聚(3-羟基丁酸酯-co-4-羟基丁酸酯)(P(3-co-4)HB)解聚酶是由微生物所分泌的一种胞外酶,可以将P(3-co-4)HB降解为水溶性小分子物质,为了阐明P(3-co-4)HB材料的生物降解机理并建立其生物循环系统,P(3-co-4)HB降解菌和降解酶的研究越来越受到关注。本论文以Penicillium sp.DS-04菌株为材料,对其产生的P(3-co-4)HB解聚酶进行了分离纯化,发酵液经过冻干和分子筛层析得到了P(3-co-4)HB解聚酶的纯酶组分,纯化倍数和回收率分别为3.15和10.44%,SDS-PAGE显示该酶分子量为44.0kDa。本文还对P(3-co-4)HB解聚酶的酶学性质进行了测定,并初步探讨了解聚酶的作用模式。     

斜卧青霉纤维素酶的研究——Ⅰ.JU_1菌株的产酶及酶作用条件

目前国内外对纤维素酶的研究多集中于木霉。但木霉毒性嫌疑大,在食品发酵工业中的应用受到限制。我们通过筛选和诱变选育,得到了一株斜卧青霉的纤维素酶抗降解物阻遏高产突变株—JU1,产酶能力近似于国内外的一些优良木霉菌株。对它的产酶条件及酶作用条件做了进一步研究。材料和方法