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.     

温度、光照对微小亚历山大藻Z1产毒含量和组成的影响

本实验在试图通过一株微小亚历山大藻Z1(AlexandriumminutumZ1)来生产和制备膝沟藻毒素(gonyautoxin GTX)的过程中,利用不同温度、光照强度对Z1进行批次培养,采用浮游植物计数板法和高效液相色谱串联质谱检测法(HPLC-MS)监测生长状况和检测毒素的含量和组成,探索适宜的生长和产毒条件。结果表明,微小亚历山大藻Z1在15~30℃的温度范围内和15.6~125μEm-2s-1光强范围内都可生长和增殖,最适生长温度为25℃,最适生长光强为62.5~125μEm-2s-1。随着培养时间的延长,毒素含量出现先增加后减少的趋势,最大值出现在平稳期,而毒素中GTX1/4占总毒素的摩尔含量比不断上升,相应地GTX2/3的占比不断下降。适中的温度(25℃)和光照强度(62.5μEm-2s-1)都可以促进Z1的毒素总含量及GTX1/4占总毒素的摩尔含量。掌握这些产毒规律,为利用微小亚历山大藻在生产麻痹性贝毒提供理论指导。     

Occurrence and Diversity of Pichia spp. in Marine Environments

A total of 328 yeast strains from seawater, sediments, mud of saltems, 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 luv-small, Pichia ohmeri YF04d, Pichiafermentans 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.     

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.     

江苏沿海贝类软体中麻痹性贝毒素种类及含量变化的多年观测研究

Paralytic shellfish toxins(PSTs) are notorious neurotoxins that threaten public health and food safety worldwide.Although PST monitoring programs have recently been established throughout China, the profiles and variation of PSTs in important commercial clams(e.g., Mactra veneriformis, Ruditapes philippinarum, and Meretrix meretrix) along the Jiangsu Province coastline remain largely unexplored. In this study, a validated hydrophilic interaction liquid chromatography–tandem mass spectrometry(HILIC-MS/MS) method was used to examine PST profiles and levels in 540 clam samples from natural production areas along Jiangsu Province coastline during2014–2016. Although the PST levels(≤6.38 μg saxitotoxin equivalents(eq)/kg) were consistently below European Union regulatory limits(≤800 μg saxitotoxin eq/kg) during this time period, saxitotoxin, decarbamoylsaxitotoxin,and gonyautoxins 1 and 4 were detected, and nearly 40% of the samples were saxitotoxin-positive. The PST levels also varied significantly by seasons, with peak values observed in May during 2014–2016. This is the first systematic report of PSTs in clams from Jiangsu Province, and additional research and protective measures are needed to ensure the safety of clams harvested in this area.     

麻痹性贝毒在毛蚶体内的转化过程研究

麻痹性贝毒能够在贝类体内累积,威胁海产品消费者健康。在以往调查中,多次在毛蚶(Scapharca subcrenata)体内发现高含量的麻痹性贝毒,但对于毛蚶体内麻痹性贝毒的转化过程及其食品安全风险还缺乏认识。通过室内模拟实验,选择太平洋亚历山大藻(Alexandrium pacificum)和链状裸甲藻(Gymnodinium catenatum)作为产毒藻种,研究了两种有毒藻种所产麻痹性贝毒在毛蚶体内的转化过程。结果表明,毛蚶体内主要出现了三种麻痹性贝毒转化过程,一是R1位羟基的还原反应,二是N-磺酰氨甲酰基类毒素R4位磺酸基团的水解反应,三是含羟基苯甲酸(hydroxybenzoate)基团的链状裸甲藻毒素在R4位的水解反应。毛蚶体内麻痹性贝毒的生物转化过程复杂,对毛蚶毒性的影响具有一定的不确定性,未来仍需要进一步深化毛蚶体内毒素累积、代谢、转化过程的研究,同时加强对毛蚶体内毒素含量的全面监测,防范毛蚶可能导致的麻痹性贝毒中毒风险。     

Effects of nitrate and phosphate on growth and C2 toxin productivity of Alexandrium tamarense CI01 in culture

Growth and C2 toxin productivity of a marine dinoflagellate, Alexandrium tamarense CI01 (ATCI01) which predominantly produces C2 toxin, were studied in unialgal batch cultures to optimize the concentrations of nitrate and phosphate for a maximal toxin yield. A range of start concentrations of the two major nutrients was determined in which algal growth was proportional to the nutrient concentrations used. The highest concentrations of nitrate and phosphate in this growth-enhancing range were 264 and 20 μM, respectively. In this concentration range, the C2 toxin yield (μg/l) and cellular toxin content (Qt, fmole per cell) reached a maximum at the lowest end of phosphate (5 μM) and the highest end of nitrate (264 μM). Further increase in the supply of nitrate continued to enhance the toxin yield. Our results indicated that the growth and toxin productivity of this algal strain in batch cultures had distinctly different optimal ranges of nitrate and phosphate concentrations. For a maximum toxin yield, a judicious use of phosphate under a nitrate-replete condition is called for.     

Effects of nitrate and phosphate on growth and C2 toxin productivity of Alexandrium tamarense CI01 in culture

Growth and C2 toxin productivity of a marine dinoflagellate, Alexandrium tamarense CI01 (ATCI01) which predominantly produces C2 toxin, were studied in unialgal batch cultures to optimize the concentrations of nitrate and phosphate for a maximal toxin yield. A range of start concentrations of the two major nutrients was determined in which algal growth was proportional to the nutrient concentrations used. The highest concentrations of nitrate and phosphate in this growth-enhancing range were 264 and 20 μM, respectively. In this concentration range, the C2 toxin yield (μg/l) and cellular toxin content (Qt, fmole per cell) reached a maximum at the lowest end of phosphate (5 μM) and the highest end of nitrate (264 μM). Further increase in the supply of nitrate continued to enhance the toxin yield. Our results indicated that the growth and toxin productivity of this algal strain in batch cultures had distinctly different optimal ranges of nitrate and phosphate concentrations. For a maximum toxin yield, a judicious use of phosphate under a nitrate-replete condition is called for.     

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 trituberculatus.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.     

两种亚历L山大藻产毒过程和毒素特征研究

对 2种有毒亚历山大藻——引自台湾的微小亚历山大藻 AMTK- ( Alexandriumminutum AMTK- )和引自美国阿拉斯加的亚历山大藻 KW0 6( Alexandrium sp.KW0 6)——在不同生长阶段的产毒状况、毒素组成和毒性大小进行了研究 ,比较了两者的异同。结果显示 ,2种藻的单位细胞毒素含量与培养阶段的变化趋势是一致的 ,即在对数生长初期达最高 ,之后逐渐下降 ;微小亚历山大藻 AMTK- 主要生产 GTXs毒素 ,从对数生长后期到静止期其毒素组成比有所变化 ,GTX1 ,GTX4比例增高而 GTX2 ,GTX3降低 ,亚历山大藻KW0 6主要生产 STX毒素 ;亚历山大藻 KW0 6的毒性高于微小亚历山大藻 AMTK- 。