Andreeva N A, Kulakovskaia T V, Kulaev I S
Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Moscow oblast, Russia.
Mikrobiologiia. 2000 Jul-Aug;69(4):499-505.
Halobacterium salinarium grown in a liquid medium consumed up to 75% of phosphates originally present in the growth medium and accumulated up to 100 mumol Pi/g wet biomass by the time it entered the growth retardation phase. The content of acid-soluble oligophosphates in the biomass was maximum at the early stage of active growth and drastically decreased when cells reached the growth-retardation phase. The total content of alkali-soluble and acid-insoluble polyphosphates changed very little throughout the cultivation period (five days). The polyphosphate content of H. salinarium cells was close to that of yeasts and eubacteria. The pyrophosphatase, polyphosphatase, and nonspecific phosphatase activities of H. salinarium cells were several times lower than those of the majority of eubacteria. The specific activity of pyrophosphatase, the most active hydrolase of H. salinarium, gradually increased during cultivation, reaching 540 mU/mg protein by the end of the cultivation period. Half of the total pyrophosphatase activity of this halobacterium was localized in the cytosol. The molecular weight of pyrophosphatase, evaluated by gel filtration, was 86 kDa. The effective Km of this enzyme with respect to pyrophosphate was 115 microM.
在液体培养基中生长的盐生盐杆菌消耗了生长培养基中高达75%的磷酸盐,并在进入生长迟缓期时积累了高达100 μmol Pi/g湿生物量。生物量中酸溶性寡磷酸盐的含量在活跃生长的早期最高,当细胞达到生长迟缓期时急剧下降。碱溶性和酸不溶性多磷酸盐的总含量在整个培养期(五天)内变化很小。盐生盐杆菌细胞的多磷酸盐含量与酵母和真细菌的相近。盐生盐杆菌细胞的焦磷酸酶、多磷酸酶和非特异性磷酸酶活性比大多数真细菌低几倍。盐生盐杆菌中最活跃的水解酶焦磷酸酶的比活性在培养过程中逐渐增加,在培养期结束时达到540 mU/mg蛋白质。这种嗜盐菌总焦磷酸酶活性的一半位于胞质溶胶中。通过凝胶过滤评估,焦磷酸酶的分子量为86 kDa。该酶对焦磷酸的有效Km为115 μM。