Phosphorylation enzymes of the propionic acid bacteria and the roles of ATP inorganic pyrophosphate, and polyphosphates.

Abstract

It is shown that polyphosphates are not generated in significant amounts in the phosphoglycerate kinase reaction; polyphosphate is more effective than ATP in the formation of glucose 6-P by glucokinase, but the rate with ATP may be adequate to meet the requirements of glucose metabolism; PP1 is far more effective than ATP as a phosphate donor in the formation of fructose 1,6-P2 by phosphofructokinase; PPj rather than ATP almost certainly is used in this reaction; and, aside from glucokinase and phosphofructokinase, the enzymes of phosphorylation are specific in their requirements of phosphate donors or acceptors and are present in adequate amounts to meet the requirements of glucose metabolism by the propionic acid bacteria.The propionic acid bacteria are unusual in that they utilize PP, in several phosphorylation reactions in place of ATP (1, 2). (5) have shown that they form poly(P) from ATP (Reaction 5).polyphosphate glucokinase glucose + poly(P)nglucose 6-P + poly(P),,1 [4] polyphosphate kinase ATP + poly(P),', ADP + poly(P),+1 [5] Of special interest is the report by Uryson and Kulaev (3) that Propionibacterium freudenreichii has poly(P) glucokinase activity but only a trace of ATP glucokinase activity.This finding raises the possibility that P. freudenreichii phosphorylates glucose almost exclusively with poly(P). In addition, Kulaev et al. (4) MATERIALS AND METHODSCultures and Conditions for Growth. Propionibacterium arabinosum, ATCC 4965; P. freudenreichii, ATCC 6207; and P. shermanii, 52W were investigated. The latter was originally isolated by Virtanen (8) and was later given the number 52W by C. H. Werkman (Iowa State University). The bacteria were grown at 30°C in 15 liters of medium containing glucose, 45 g; Na2CO2, 82.5 g; KH2PO4, 153 g; yeast extract, 76 g; Co(NO3)266H2O, 0.15 g; thiamin HCl, 15 mg; calcium pantothenate, 15 mg; biotin, 7.5 mg; and 20 ml of trace elements (boric acid, 75 mg; CuS04, 6 mg; Nal, 13.5 mg, FeSO4, 66.2 mg; ZnCl2, 28.4 mg; and SeO2, 16.6 mg per 100 ml). Glucose and Na2CO3 were each sterilized separately. Glycerol medium was identical except 450 g of glycerol was substituted for glucose. Lactate medium contained (per 15 liters) sodium lactate (USP 60% syrup, Pfansted Laboratories, Worthington, IL), 750 ml; yeast extract, 60 g; tryptone, 30 g; K2HPO4, 60 g; NaH2PO4, 30 g; FeSO4, 0.15 g; and CoNO3, 0.15 g; vitamins and trace elements were as given above for the glucose medium. Bacteria were harvested using a Sharples centrifuge, either after 4-6 days during the stationary phase of growth (Table 1) or after 2, 4, and 7 days of growth (Table 2). A portion of the 15 liters of culture was removed and the gas phase was restored with CO2 after each removal to maintain anaerobic conditions. In the experiments of Tables 1 and 2, the packed cells were held at -70°C until preparation of the crude extract.Crude Extracts and Fractionations. The cells were broken by passage through a French press or by grinding in an Ep- 312The publication costs of this article were defray...

Physiology of the wall-less Anaeroplasmataceae (Class Mollicutes) and related walled bacteria

“…acidlpropionici, some Rhodopseudomonas spp., Brevibacterlum amnonlagenes i"Corynebacterlum awmonlagenes"), and the Clostridium isolated from the cockroach gut Wood and Goss, 1985;Wood et al, 1969). Acholeplasm laldlawll was reported to possess activity for PEP carboxytransphosphorylase , but more recently, workers from the same laboratory have not been able to confirm this activity .…”

“…Acetobacter spp., the Clostridium isolated from the cockroach gut, and three anoxygenic photosynthetic bacteria Wood and Goss, 1985). Among the mollicutes, PPDK has been detected in As.…”

“…arabinosum") Wood and Goss, 1985), some Bacteroides spp. (Roberton and Qluclna, 1982), and Deleya spp.…”

“…, Euglena gracilis (Miyatake et al, 1984), Propionibacterium spp. Wood and Goss, 1985), and the Bacteroides spp. (Roberton and Gluclna, 1982) have been reported to also have ATP-PFK, but Deleya spp.…”

“…Pollack (1986) examined U. urealytlcum for glycolytic enzymes; hexokinase, Glc-6-P Wood and Goss, 1985), some Bacteroldes spp. (Hacy et al, 1978;Roberton and Glucina, 1982), Deleya spp.…”