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嗜冷菌雪腐单端孢霉在限定培养基中的深层分批培养;生长、碳水化合物利用及对温度的响应

Submerged batch culture of the psychrophile Monographella nivalis in a defined medium; growth, carbohydrate utilization and responses to temperature.

作者信息

Cairns Andrew J, Howarth Catherine J, Pollock Christopher J

机构信息

Cell Biology, Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, SY23 3EB, UK.

Environmental Biology Departments, Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, SY23 3EB, UK.

出版信息

New Phytol. 1995 Feb;129(2):299-308. doi: 10.1111/j.1469-8137.1995.tb04300.x.

DOI:10.1111/j.1469-8137.1995.tb04300.x
PMID:33874546
Abstract

An asporogenous strain of the pink snow mould fungus, Monographella nivalis (Schaffnit) E. Müller, anamorph Gerlachia nivalis (Ces. ex Sac.) W. Gams & E. Müller (Syn. Fusarium nivale Ces. ex Sacc.), grew at 5 °C on a denned salts medium plus vitamins and utilized a variety of simple and polymeric carbohydrates as the sole carbon and energy source. Mycelium was grown at temperatures between 3 and 15 °C in aerated submerged fermentation culture in chemically defined medium plus sucrose. Optimum growth rates of 0·035-0·033 h occurred between 9 and 12 °C. Growth in a simple medium showed that all biochemical and physiological processes necessary for growth were functional at 3 °C. The growth performance of the organism at low temperatures was no better than would be expected from extrapolation of mesophilic growth responses to temperature. The optimum growth temperature of 9-12°C showed that some biochemical or physiological process was impaired above 12 °C. Uptake and incorporation of S-methionine by mycelium at different temperatures showed that general protein synthesis increased up to 25 °C, and hence was not responsible per se for the sensitivity to temperatures above 12 °C. Heat shock proteins were synthesized at the relatively low temperature of 25 °C, consistent with the low temperature optimum for growth. When grown with sucrose as the sole carbon source, the mycelium catalyzed the extracellular hydrolysis of sucrose, releasing glucose and fructose together with a small amount of fructan trisaccharides and a trace of tetra- and penta-saccharides. Fructan accumulation was transient, corresponding with maximal rates of sucrose hydrolysis. Most biomass formation occurred in the absence of fructan in the culture, hence fructan was not necessary for growth at low temperature and did not appear to function as a cryoprotectant. Invertase activity was mostly (60-70%) bound to mycelium; the remainder was free in the culture supernatant. The regulation of invertase expression appeared to be by sucrose-induction, rather than by end-product repression. Rates of sucrose hydrolysis in culture were temperature-sensitive and were markedly depressed above 12 °C, indicating inhibition of invertase formation.

摘要

粉红雪霉病菌(Monographella nivalis (Schaffnit) E. Müller,无性型为Gerlachia nivalis (Ces. ex Sac.) W. Gams & E. Müller,同义词为Fusarium nivale Ces. ex Sacc.)的一个不产孢菌株,能在5°C的限定盐类培养基加维生素的条件下生长,并利用多种简单和聚合碳水化合物作为唯一的碳源和能源。菌丝体在3至15°C的温度下,于通气的深层发酵培养中,在化学限定培养基加蔗糖的条件下生长。最佳生长速率为0.035 - 0.033 h⁻¹,出现在9至12°C之间。在简单培养基中的生长表明,生长所需的所有生化和生理过程在3°C时都能正常运作。该生物体在低温下的生长表现并不比根据嗜温生长对温度的响应进行外推所预期的更好。9至12°C的最佳生长温度表明,某些生化或生理过程在12°C以上会受到损害。不同温度下菌丝体对³⁵S - 蛋氨酸的摄取和掺入表明,一般蛋白质合成在25°C时达到最高,因此本身并非对12°C以上温度敏感的原因。热休克蛋白在相对较低的25°C温度下合成,这与生长的低温最佳温度一致。当以蔗糖作为唯一碳源生长时,菌丝体催化蔗糖的细胞外水解,释放出葡萄糖和果糖,同时还有少量的果聚糖三糖以及微量的四糖和五糖。果聚糖的积累是短暂的,与蔗糖水解的最大速率相对应。大多数生物量的形成发生在培养物中不存在果聚糖的情况下,因此果聚糖对于低温生长并非必需,而且似乎也不起抗冻保护剂的作用。转化酶活性大部分(60 - 70%)与菌丝体结合;其余部分游离于培养上清液中。转化酶表达的调控似乎是由蔗糖诱导,而非终产物阻遏。培养物中蔗糖水解速率对温度敏感,在12°C以上明显降低,表明转化酶的形成受到抑制。

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