A 31P NMR study of the internal pH of yeast peroxisomes

The internal pH of peroxisomes in the yeasts Hansenula polymorpha, Candida utilis and Trichosporon cutaneum X4 was estimated by ³¹P nuclear magnetic resonance (NMR) spectroscopy. ³¹P NMR spectra of suspensions of intact cells of these yeasts, grown under conditions of extensive peroxisomal proliferation, displayed two prominent P_i-peaks at different chemical shift positions. In control cells grown on glucose, which contain very few peroxisomes, only a single peak was observed. This latter peak, which was detected under all growth conditions, was assigned to cytosolic P_i at pH 7.1. The additional peak present in spectra of peroxisome-containing cells, reflected P_i at a considerably lower pH of approximately 5.8–6.0. Experiments with the protonophore carbonyl cyanide m-chlorophenylhydrazon (CCCP) and the ionophores valinomycin and nigericin revealed that separation of the two P_i-peaks was caused by a pH-gradient across a membrane separating the two pools. Experiments with chloroquine confirmed the acidic nature of one of these pools. In a number of transfer experiments with the yeast H. polymorpha it was shown that the relative intensity of the P_i-signal at the low pH-position was correlated to the peroxisomal volume fraction. These results strongly suggest that this peak has to be assigned to P_i in peroxisomes, which therefore are acidic in nature. The presence of peroxisome-associated P_i was confirmed cytochemically.Abbreviations CCCP Carbonyl cyanide m-chlorophenylhydrazon - DCCD N,N-dicyclohexylcarbodiimide     

Oxidation of amines by yeasts grown on 1-aminoalkanes or putrescine as the sole source of carbon,nitrogen and energy

The maximum growth rate of Trichosporon cutaneum CBS 8111 in chemostat cultures was 0.185 h^-1 on ethylamine and 0.21 h^-1 on butylamine, that of Candida famata CBS 8109 was 0.32 h^-1 on putrescine.The amine oxidation pattern of the ascomycetous strains studied, viz. Candida famata CBS 8109, Stephanoascus ciferrii CBS 4856 and Trichosporon adeninovorans CBS 8244 was independent of the amine that had been used as the growth substrate. It resembled that of benzylamine/putrescine oxidase found in other ascomycetous yeasts. However, differences in pH optimum and substrate specificity were observed between the amine-oxidizing systems of these three species.The amine oxidation pattern of cell-free extracts of Trichosporon cutaneum CBS 8111 varied with the amine that was used as growth substrate. The enzyme system produced by Cryptococcus laurentii CBS 7140 failed to oxidize isobutylamine and benzylamine, and showed a high pH optimum.The synthesis of amine oxidase in the four yeast strains studied was not repressed by ammonium chloride and was weakly repressed by glucose but was strongly repressed if both compounds were present in the growth medium.     

双孢蘑菇上的丝枝霉新种

1980—1981年先后自北京、昆明、福州等地双孢蘑菇[Agaricus bisporus (Lange)Sing.]上分离到一种真菌,在PDA上的培养性状与蘑菇轮枝菌(Vertieillium psalliotae Tresch．)和蛛网丝枝霉中国变种[Aphanocladium aranearum(Petch) Gams var sincase J．D．Chen]近 似。镜检可见其兼具上述两种菌的形态。经反复多次纯化,并挑取单孢子培养,肯定了这种真菌是纯菌。在培养基上已保存4年,形态稳定。它具典型蘑菇轮枝菌形态,而其丝枝霉型瓶梗的形成和瘪缩时间、长度、原瓶梗数量、末端瓶梗数及排列密度等,都与蛛网丝枝霉中国变种有一定差异,与丝枝霉属其它种亦不同,故定为两型丝枝霉新种(Aphanocladium dimorphum sp-nov.)。本文详细描述了该新种的形态,井对文甲所提的五种真菌及其与Engyodontium属间的关系进行了讨论。     

丝孢酵母高甲硫氨酸突变株的选育及营养调控

以丝孢酵母(Trichosporon Behr)ST851为原始菌株,经紫外线诱变,在含乙硫氨酸的双层平板上筛选到多株抗乙硫氨酸突变株。其中ST851-10株抗乙硫氨酸浓度达到350μg/ml,其菌体蛋白质含量由40.5％提高到44.3％,菌体甲硫氨酸含量由20.45mg/g-DCW增加到29.32mg/g-DCW。在以苹果渣为碳源、尿素为氮源、硫酸镁作硫源的最适培养条件下,固态发酵24h后,蛋白质和甲硫氨酸含量较原始菌株分别提高了15.8％和44.9％。培养基中C/N值低有利于甲硫氨酸的合成,C/N值高则适合于菌体生长。在苹果渣固态发酵过程中,适当补加氮源既有利于菌体生长和甲硫氨酸的合成,又可起到调节培养基pH值的作用。     

不同钾水平对钾饥饿墨兰碳水化合物和蛋白质含量的影响

墨兰（Ｃｙｍｂｉｄｉｕｍｓｉｎｅｎｓｅ（Ａｎｄｒ．）Ｗｉｌｌｄ）植株经过钾饥饿后,无上栽培于不同钾浓度的培养液中．随着钾浓度的升高（５ｍｍｏｌ／Ｌ）,体内可溶性糖、淀粉、纤维素和蛋白质含量比对照分别增加１２５、１１７、１２７和４１％,而还原糖和游离氨基酸含量则比对照分别下降４４％和２４％．假球茎是贮藏还原糖、可溶性糖、淀粉、游离氨基酸和蛋白质的主要器官,叶片是纤维素最多的器官．钾供应充足时,叶片丙酮酸激酶活性明显加强（比对照强１５倍）,而硝酸还原酶活性也加强（比对照强０．８倍）．本文对钾促进墨兰生长发育和抗病等原因加以讨论．并初步提出诊断墨兰体内钾状况的三种生理指标．     

Biotransformation of diphenyl ether by the yeastTrichosporon beigelii SBUG 752

Trichosporon beigelii SBUG 752 was able to transform diphenyl ether. By TLC, HPLC, GC, GC-MS, NMR- and UV-spectroscopy, several oxidation products were identified. The primary attack was initiated by a monooxygenation step, resulting in the formation of 4-hydroxydiphenyl ether, 2-hydroxydiphenyl ether and 3-hydroxydiphenyl ether (48:47:5). Further oxidation led to 3,4-dihydroxydiphenyl ether. As a characteristic product resulting from the cleavage of an aromatic ring, the lactone of 2-hydroxy-4-phenoxymuconic acid was identified. The possible mechanism of ring cleavage to yield this metabolite is discussed.     

寄生双孢蘑菇的蛛网丝枝霉中国变种

1980—1981年自北京、福州、三明、泉州、漳州、厦门和昆明等地双孢蘑菇[Agaricusbisporus(Lange)Sing]病菇上和不出蘑菇的培养料内,分离到一种真菌。在 PDA 培养基上的培养性状,肉眼观察与蘑菇轮枝菌(Verticillium psalliotae Tresch.)极难区别。根据其分生孢子和瓶梗的形态及其在匐匍气丝上的排列情况等,鉴定为珠网丝枝霉。因具短棒状原瓶梗,认为是一新变种,定名为珠网丝枝霉中国变种 Aphanocladium aranearum(Petch)W.Gams var.sinense J.D.Chen var.nov.它是双孢蘑菇的重要害菌之一。这个“属”和“种”都是我国的新记录。在猴头(Hericium erinaceus)、香菇(Lentinus edo-des),糙皮侧耳(Pleurotus ostreatus)上和水稻土中也有少量分布。     

粘性丝孢酵母的诱变及以玉米秸秆水解液为原料产油条件优化

对粘性丝胞酵母进行紫外诱变,获得一株产油率较高的菌株,较原菌株提高了1.53倍。将该菌株接种于用1%硫酸和酶水解处理并浓缩至还原糖浓度为5%的玉米秸秆水解液中培养,生长较好。通过四因素三水平正交实验,确定培养条件为初始pH值7.0、接种量1%、发酵温度30℃、发酵时间5 d时产油率最高。对最佳产油条件进行验证,测得油脂含量为21.3%。从而为利用农业废弃物大规模生产微生物油脂提供了试验数据。气相色谱-质谱联用分析仪显示油脂脂肪酸组成为棕榈酸28.36%,油酸55.86%,10-十八烯酸9.23%,硬脂酸6.70%,可以作为原料生产生物柴油。     

翼蓼和中华抱茎蓼挥发油化学成分分析

采用水蒸气蒸馏法分别从翼蓼和中华抱茎蓼中提取其挥发油,并用气相色谱-质谱联用技术(GC-MS)对挥发油中的化学成分进行分离和结构鉴定,运用气相色谱峰面积归一化法确定各成分的相对百分含量.结果表明:翼蓼中总共鉴定出31种成分,中华抱茎蓼中鉴定出21种成分,分别占到挥发油总含量的84.0%和93.8%,翼蓼挥发油中主要成分是表蓝桉醇(15.9%)、甲苯(8.6%)、反式橙花叔醇(6.9%);中华抱茎蓼挥发油主要成分为邻苯二甲酸二异丁酯(19.9%)、2,4-戊二酮(17.2%)、邻苯二甲酸二丁酯(11.7%)、3-甲基-2,3-二氢苯并呋喃(10.7%).     

荷叶铁线蕨孢子的不同贮藏时间与培养条件对配子体和孢子体形成的影响

研究了荷叶铁线蕨(Adiantum reniforme var.sinense)的不同贮藏时间与培养条件对其配子体发育和孢子体形成的影响,结果表明:荷叶铁线蕨孢子为不含叶绿素孢子,在室温下保存5年以上时间依然具有活力,但不同的贮藏时间对配子体和孢子体的形成具有显著的影响。其中,室温贮藏7d的配子体形成的时间为29d,而贮藏5年的为38d;贮藏7d的孢子体形成的时间为96d,贮藏4个月的为105d。在培养室[(25±1)℃恒温和每天2000~2500lx12h光照]沙基质中,从孢子播种到形成孢子体时间最短,为87d;沙基质比腐殖土更利于孢子体的形成;配子体密度在不超过3株/cm2的情况下,密度越高越有利于孢子体的形成。