Isolation and identification of a marine killer yeast strain YF07b and cloning of the gene encoding killer toxin from the yeast

It was found that the marine yeast strain YF07b could secrete a large amount of killer toxin against a pathogenic yeast strain WCY which could cause milky disease in Portunus trituberculcttus. The marine yeast strain YF07b was identified to be Pichia anomala according to the results of routine yeast identification and 18S rDNA and ITS sequences. The gene encoding killer toxin in the marine yeast strain YF07b was amplified by PCR technology. After sequencing, the results show that an open reading frame, consisting of 1 281 bp, encoded a presumed protein of 427 amino acids. The sequence of the cloned gene was found to have 99% match with that of the gene encoding killer toxin in Pichia anomalas strain K. A signal peptide including 17 amino acids appeared in the N-terminal domain of the killer toxin. Therefore, the mature protein consisted of 410 amino acids, its molecular mass was estimated to be 47.4 ku and its isoelctronic point was 4.5.

Characteristics of Killer Toxin of the Yeast <i>Cryptococcus pinus</i>

This work was aimed to study of fungicidal activity and mechanism of action of the killer toxin of Cryptococcus pinus VKM Y-2958. A killer-toxin preparation was obtained and partly purified from the culture liquid of the yeast Cryptococcus pinus. Its fungicidal activity against the pathogenic yeast Filobasidiella neoformans was assessed by assay of target cells growth inhibition and propidium iodide staining. The preparation contained three peptides with molecular masses of about 11 - 13 kDa, which did not show similarities in the mass spectrometric analysis. When frozen, the killer-toxin preparation remained active for several years. The killer toxin of Cr. pinus exhibits the high fungicidal activity against the known pathogen F. neoformans, is stable during long-term storage, and its mechanism of action is based on damaging target cell membranes. The killer toxin was obtained by a simple procedure and is of interest for the development of novel fungicidal preparations.

海洋酵母菌胞外酶及其基因的最新研究进展

综述海洋酵母菌所产多种胞外酶的生产、纯酶性质及基因克隆工作的最新研究进展。从实验室建立的500多株海洋酵母菌种资源库中筛选出了数十株可以产生多种不同活性胞外酶的海洋酵母菌,它们分别属于耶鲁维亚酵母菌(Yarrowia)、类酵母(Aureobasidium)、毕赤氏酵母菌(Pichia)、梅奇酵母菌(Metschnikowia)和隐球酵母菌(Cryptococcus)等不同种属。所产生的胞外酶主要包括纤维素酶、碱性蛋白酶、酸性蛋白酶、淀粉酶(包括生淀粉酶)、菊糖酶、脂酶、植酸酶和嗜杀因子等。深入进行了各种胞外酶的生产,纯化及纯酶的性质研究,并对部分菌株的胞外酶基因进行了克隆、测序和表达,这些研究有助于对海洋酵母胞外酶的理论研究和生产应用价值的了解和对于海洋酵母菌资源、生理和遗传的进一步深入研究。

嗜杀酵母的生物学功能及其应用

嗜杀酵母分泌的外毒素可以抑制或杀死敏感菌株,并且对自身分泌的嗜杀因子具有免疫性。将嗜杀酵母用于工业生产中,可以净化环境,提高产品的质量;一些海洋嗜杀酵母还可以用于海水养殖业,可以控制某些海洋酵母菌引起的海洋动物疾病。除此之外,嗜杀酵母分泌的毒素蛋白可以作为潜在的抗真菌药物,以便抵制某些病原酵母菌或类酵母菌等微生物引起的侵染。介绍嗜杀酵母的种类、作用机理,并且概述嗜杀酵母的应用及研究进展。

野生嗜杀白假丝酵母LFA418的产毒条件优化及其毒素粗提物特性

对1株分离自新疆楼兰葡萄酒厂野生白假丝酵母(Candida albicans)LFA418的嗜杀特征及其粗毒素特性进行研究。结果显示,C.albicans LFA418具有致死克鲁维毕赤酵母(Pichia kluyveri)、贝酵母(Saccharomyces bayanus)、Lachancea thermotolerans和葡萄牙假丝酵母(Candida lusitaniae)的嗜杀性。接种体积分数7%C.albicans LFA418到YEPD液体培养基中,25℃培养32 h,可以获得最大产量的嗜杀毒素;利用截留分子质量为8 000 D的透析袋制备的粗毒素,在温度分别-20、4℃时pH 4.2~4.6和pH 4.2时温度4~20℃具有嗜杀活性的稳定性。

嗜杀酵母的研究进展及其在酿酒工业中的应用

嗜杀酵母能够分泌毒素蛋白抑制或杀死敏感细胞,在发酵生产中利用其特性可以净化环境,提高产品的质量。文中介绍了嗜杀酵母的分布、作用机理以及嗜杀酵母质粒的提取和选育,并概述了嗜杀酵母在酿酒工业中的应用和研究进展。

响应面法优化拟威尔嗜杀酵母Cyberlindnera mrakii WM1代谢产嗜杀因子的发酵条件

以保藏于本实验室的海洋酵母菌种资源库的酵母菌株Cyberlindnera mrakii WM1为研究对象,以人类条件致病菌白色假丝酵母Canidia albicans YTS-03为嗜杀毒素抑菌活性测试指示菌,以抑菌圈直径为考察指标,对拟威尔嗜杀酵母WM1代谢产嗜杀因子的发酵条件进行优化。首先以接种量、发酵液p H、发酵温度和时间为自变量,确定其四个因素对嗜杀活性的抑菌效果影响显著,再对这四个因素进行响应面实验优化。获得WM1生长代谢产嗜杀因子的最佳发酵条件为:接种量8%,发酵液p H4.0,发酵温度21℃,发酵时间2.5 d。优化后的WM1酵母发酵液的抑菌圈的直径达到21.00 mm,比优化前(16.5 mm)提高27.3%;菌液浓度也达2.58×10~8CFU/m L,比优化前(1.9×10~8CFU/m L)提高了35.8%。同时,最优发酵条件下获得的实验结果与模型预测值相吻合,说明建立的回归模型是切实可行的。

嗜杀酿酒酵母毒素蛋白及其杀伤质粒的研究

Killer toxin from \%Saccharomyces cerevisiae\% SK was isolated by ultrafiltration of culture supernatants and purified by poly(ethylene glycol). The toxin migrates as one single protein band on SDS-PAGE and its molecular weight is 15kD. The SK toxin has the greatest lethal effect on the sensitive yeast strain in the lat lag phase. Extraction and purification of killer heretity factor(dsRNA) from SK found that M dsRNA plasmid and L dsRNA plasmid have different molecular lengths being 1.7kb and 4.0kb.

嗜杀酵母的生物学功能及其应用

嗜杀酵母能够分泌毒素蛋白,杀死敏感酵母。嗜杀酵母对自身分泌的嗜杀毒素具有免疫力。嗜杀酵母的嗜杀特性与两种双链线状RNA(dsRNA)有关,即编码产生毒素蛋白的M-dsRNA和编码自身和M-dsRNA外壳蛋白的L-dsRNA。嗜杀毒素破坏细胞跨膜化学质子梯度,造成ATP和钾离子泄漏,导致细胞死亡。应用嗜杀酵母可避免野生型酵母污染,净化发酵体系,改善发酵产物品质;嗜杀毒素也可作为抵制病原酵母和类酵母微生物的抗真菌剂。

Occurrence and Diversity of Marine Yeasts in Antarctica Environments

A total of 28 yeast strains were obtained from the sea sediment of Antarctica.According to the results of routine identi-fication and molecular characterization,the strains belonged to species of Yarrowia lipolytica,Debaryomyces hansenii,Rhodotorula slooffiae,Rhodotorula mucilaginosa,Sporidiobolus salmonicolor,Aureobasidium pullulans,Mrakia frigida and Guehomyces pullu-lans,respectively.The Antarctica yeasts have wide potential applications in biotechnology,for some of them can produce b-galactosidase and killer toxins.

嗜杀酵母的生物学特性及应用展望

嗜杀酵母能够分泌嗜杀毒素杀死特定的微生物。将嗜杀酵母作为工业生产用菌株,可以有效防止生产过程中的杂菌污染,净化发酵体系,保证发酵的正常进行;嗜杀毒素还可以制备成为抗真菌剂,用以抵制病原酵母及类酵母等微生物的侵染。综合论述了嗜杀酵母的分类学地位及生物学特性,嗜杀毒素产生和作用的机理,并阐述了环境因素对嗜杀酵母菌自身及其分泌嗜杀毒素的影响。

Intraspecific Diversity of Aureobasidium pullulans Strains from Different Marine Environments

Totally more than 500 yeast strains were isolated from seawater, sea sediments, mud of sea salterns, marine fish guts and marine algae. The results of routine and molecular biology identification methods show that nine strains among these marine yeasts belong to Aureobasidium pullulans, although the morphologies of their colonies are very different. The marine yeasts isolated from different marine environments indicate that A. pullulans is widely distributed in different environmental conditions. These Aureo-basidium pullulans strains include A. pullulans 4#2, A. pullulans N13d, A. pullulans HN3-11, A. pullulans HN2-3, A. pullulans JHSc, A. pullulans HN4.7, A. pullulans HN5.3, A. pullulans HN6.2 and A. pullulans W13a. A. pullulans 4#2 could produce cellulase and single cell protein. A. pullulans N13d could produce protease, lipase, amylase and cellulase. Both A. pullulans HN3-11 and A. pullulans HN2-3 were able to produce protease, lipase and cellulase. A. pullulans JHSc could secrete cellulase and killer toxin. Both A. pullulans HN4.7 and A. pullulans HN5.3 could yield lipase and cellulase. A. pullulans W13a was able to secrete extracellular amylase and cellulase while A. pullulans HN4.7 and A. pullulans N13d could produce siderophores. This means that different A. pullulans strains from different marine environments have different physiological characteristics, which may be applied in many different biotechnological industries.

酵母杀手毒素的作用机制及应用研究进展

杀手毒素的作用机制与抑制酵母细胞壁 β 葡聚糖合成有关。甘露糖蛋白可能是HM 1的受体之一 ,其抗性的产生是N 糖基化减少所致。HM 1是 88个氨基酸组成的亲水蛋白质 ,C 末端 82和 86位精氨酸对其杀手活性至关重要。

酿酒酵母两种不同嗜杀菌株的比较研究

以酿酒酵母两种不同类型的嗜杀菌株SK4（K1型）和ERRI（K2型）为材料，分析了不同嗜杀酵母的嗜杀特性，两株嗜杀酵母具有相互杀死作用，其嗜杀活性与菌体生长有关。SK4和ERRI的嗜杀质粒的比较表明：M1－dsRNA质粒和M2-dsRNA质粒分子量分别为1．7kb和1．5kb，两株菌的L－dsRNA质粒均为4．0kb。用高温和紫外线处理嗜杀酵母，嗜杀活性随之消失，消除菌中的M－dsRNA质粒也相应消失，嗜杀活性的消除率随菌株和消除剂的不同而变化。实验证明两株菌产生的毒素蛋白的最适嗜杀作用条件不同，最适pH和温度分别为4．8、16℃和4．0、22℃，但两种毒素蛋白均对在对数生长期的敏感细胞作用最显著。

Occurrence and Diversity of Pichia spp. in Marine Environments

A total of 328 yeast strains from seawater, sediments, mud of salterns, the guts of marine fish and marine algae were obtained. The results of routine identification and molecular methods show that five yeast strains obtained in this study belonged to Pichia spp., including Pichia guilliermondii 1 uv-small, Pichia ohmeri YF04d, Pichia fermentans YF12b, Pichia burtonii YF11A and Pichia anomala YF07b. Further studies revealed that Pichia anomala YF07b could produce killer toxin against pathogenic yeasts in crabs while Pichia guilliermondii luv-small could produce high activity of extracellular inulinase. It is advisable to test if Pichia ohmeri YF04d obtained in this study is related to central-venous-catheter-associated infection.

嗜杀酵母的生物学特性及其应用

嗜杀酵母在生长繁殖过程当中所产的嗜杀因子能够抑制或杀死某些特定的微生物,而这种毒素对于酵母本身却没有嗜杀作用。将嗜杀酵母应用于工业生产中,可以抑制野生酵母的污染、净化空气、提高产品质量,在发酵当中对体系环境的净化和生物稳定性具有较好的效果;一些海洋嗜杀酵母因能杀灭某些海洋动物的致病菌,而被应用于海水养殖业;嗜杀酵母分泌的嗜杀因子可作为抑制某些病原酵母及类酵母的抗真菌药制剂;此外,嗜杀酵母在基因工程和转基因植物当中也有应用,用作为研究酵母菌分子生物学的良好基因工程载体以及大量合成和分泌嗜杀因子以对抗病原真菌的侵染。本文就嗜杀酵母的种类、生物特性及嗜杀因子作用机理进行了综合论述,概述了嗜杀酵母在不同领域的应用,并对其应用前景进行了展望。

翁连解毒汤治疗浊毒内蕴型溃疡性结肠炎的疗效观察

目的:探究翁连解毒汤治疗浊毒内蕴型溃疡性结肠炎(Ulcerative Colitis,UC)临床疗效及可能的作用机制。方法:选取2016年2月至2018年12月在攀枝花学院附属医院收治的浊毒内蕴型UC患者130例作为研究对象,按照入院先后顺序分为对照组和观察组,每组65例。对照组常规西医治疗,观察组加用翁连解毒汤,观察2组治疗前、完成治疗后T细胞、NK细胞、炎性反应因子、凝血功能、生命质量变化,完成治疗后总结疗效。结果:观察组患者在NK细胞、T淋巴细胞组成、CR、TNF-α、IL-1β、ESR、内毒素、IL-10、FIB、PLT、APTT改善方面的效果较对照组更优,组间差异有统计学意义(P<0.05)。观察组患者完成治疗后的生命质量量表各维度评分及总评分改善效果优于对照组患者(P<0.05),且临床总有效率和组织病理学疗效率均高于对照组患者(P<0.05)。结论:翁连解毒汤能抑制浊毒内蕴型UC炎性反应,改善高凝状态,提高机体免疫功能,从而提高疗效和生命质量。