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弯曲热单孢菌中纤维素酶生物合成诱导和阻遏过程中的环磷酸腺苷水平

Cyclic AMP levels during induction and repression of cellulase biosynthesis in Thermomonospora curvata.

作者信息

Wood W E, Neubauer D G, Stutzenberger F J

出版信息

J Bacteriol. 1984 Dec;160(3):1047-54. doi: 10.1128/jb.160.3.1047-1054.1984.

Abstract

Specific cellulase production rates (SCPR) were compared with intracellular cyclic AMP (cAMP) levels in the thermophilic actinomycete, Thermomonospora curvata, during growth on several carbon sources in a chemically defined medium. SCPR and cAMP levels were 0.03 U (endoglucanase [EG] units) and 2 pmol per mg of dry cells, respectively, during exponential growth on glucose. These values increased to about 6 and 25, respectively, during growth on cellulose. Detectable EG production ceased when cAMP levels dropped below 10. Cellobiose (usually considered to be a cellulase inducer) caused a sharp decrease in cAMP levels and repressed EG production when added to cellulose-grown cultures. 2-deoxy-D-glucose, although nonmetabolizable in T. curvata, depressed cAMP to levels observed with glucose, but unlike glucose, the 2DG effect persisted until cells were washed and transferred to fresh medium. SCPR values and cAMP levels in cells grown in continuous culture under conditions of cellobiose limitation were markedly influenced by dilution rate (D). The maxima for both occurred at D = 0.085 (culture generation time of 11.8 h). When D was held constant and cellobiose concentration was increased over a 14-fold range to support higher steady state population levels, SCPR values decreased about fivefold, indicating that extracellular catabolite accumulation may be a factor in EG repression. The role of cAMP in the mechanism of this repression appears to be neither simple nor direct, since large changes (up to 200-fold) in SCPR accompany relatively small changes (10-fold) in cellular cAMP levels.

摘要

在化学成分确定的培养基中,嗜热放线菌弯曲高温单孢菌在几种碳源上生长期间,比较了其特定纤维素酶产生速率(SCPR)与细胞内环磷酸腺苷(cAMP)水平。在以葡萄糖为碳源指数生长期间,SCPR和cAMP水平分别为0.03 U(内切葡聚糖酶[EG]单位)和每毫克干细胞2 pmol。在以纤维素为碳源生长期间,这些值分别增加到约6和25。当cAMP水平降至10以下时,可检测到的EG产生停止。纤维二糖(通常被认为是纤维素酶诱导剂)添加到以纤维素为碳源生长的培养物中时,会导致cAMP水平急剧下降并抑制EG产生。2-脱氧-D-葡萄糖虽然在弯曲高温单孢菌中不可代谢,但可将cAMP降低至葡萄糖培养时观察到的水平,但与葡萄糖不同的是,2-脱氧-D-葡萄糖的影响一直持续到细胞被洗涤并转移到新鲜培养基中。在纤维二糖限制条件下连续培养的细胞中,SCPR值和cAMP水平受稀释率(D)的显著影响。两者的最大值均出现在D = 0.085(培养世代时间为11.8小时)时。当D保持恒定时,将纤维二糖浓度在14倍范围内增加以支持更高的稳态群体水平,SCPR值下降约五倍,这表明细胞外分解代谢产物的积累可能是EG抑制的一个因素。cAMP在这种抑制机制中的作用似乎既不简单也不直接,因为SCPR的大幅变化(高达200倍)伴随着细胞内cAMP水平的相对较小变化(10倍)。

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