Nurminen T, Oura E, Suomalainen H
Biochem J. 1970 Jan;116(1):61-9. doi: 10.1042/bj1160061.
A study was made of the enzyme content of the isolated cell walls and of a plasma-membrane preparation obtained by centrifugation after enzymic digestion of the cell walls of baker's yeast. The isolated cell walls showed no hexokinase, alkaline phosphatase, esterase or NADH oxidase activity. It was concluded that these enzymes exist only in the interior of the cell. Further, only a negligible activity of deamidase was detectable in the cell walls. Noticeable amounts of saccharase, phosphatases hydrolysing p-nitrophenyl phosphate, ATP, ADP, thiamin pyrophosphate and PP(i), with optimum activity at pH3-4, and an activity of Mg(2+)-dependent adenosine triphosphatase at neutral pH, were found in the isolated cell walls. During enzymic digestion, the other activities appearing in the cell walls were mostly released into the medium, but the bulk of the Mg(2+)-dependent adenosine triphosphatase remained in the plasma-membrane preparation. Accordingly, it may be assumed that the enzymes released into the medium during digestion are located in the cell wall outside the plasma membrane, whereas the Mg(2+)-dependent adenosine triphosphatase is an enzyme of the plasma membrane. This enzyme differs from the phosphatases with pH optima in the range pH3-4 with regard to location, pH optimum, substrate specificity and different requirement of activators.
对分离得到的细胞壁的酶含量以及通过对面包酵母细胞壁进行酶解消化后离心获得的质膜制剂进行了研究。分离得到的细胞壁未显示己糖激酶、碱性磷酸酶、酯酶或NADH氧化酶活性。得出的结论是,这些酶仅存在于细胞内部。此外,在细胞壁中仅检测到可忽略不计的脱酰胺酶活性。在分离得到的细胞壁中发现了大量的蔗糖酶、水解对硝基苯磷酸、ATP、ADP、硫胺素焦磷酸和PP(i)的磷酸酶,其在pH3 - 4时具有最佳活性,以及在中性pH下具有Mg(2 +)依赖性腺苷三磷酸酶活性。在酶解消化过程中,细胞壁中出现的其他活性大多释放到培养基中,但大部分Mg(2 +)依赖性腺苷三磷酸酶仍保留在质膜制剂中。因此,可以假定在消化过程中释放到培养基中的酶位于质膜外的细胞壁中,而Mg(2 +)依赖性腺苷三磷酸酶是质膜的一种酶。这种酶在位置、最适pH、底物特异性和对激活剂的不同需求方面与最适pH在pH3 - 4范围内的磷酸酶不同。