Purification and kinetics of a raw starch-hydrolyzing,thermostable, and neutral glucoamylase of the thermophilic mold Thermomucor indicae-seudaticae

The purified glucoamylase of the thermophilic mold Thermomucor indicae-seudaticaehad a molecular mass of 42 kDa with a pI of 8.2. It is a glycoprotein with 9-10.5% carbohydrate content, which acted optimally at 60 degrees C and pH 7.0, with a t(1/2) of 12 h at 60 degrees C and 7 h at 80 degrees C. Its experimental activation energy was 43 KJ mol(-1) with temperature quotient (Q(10)) of 1.35, while the values predicted by response surface methodology (RSM) were 43 KJ mol(-1) and 1.28, respectively. The enzyme hydrolyzed soluble starch at 50 degrees C (K(m) 0.50 mg mL(-1) and V(max) 109 micromol mg(-1) protein min(-1)) and at 60 degrees C (K(m) 0.40 and V(max) 143 micromol mg(-1) protein min(-1)). The experimental K(m) and V(max) values are in agreement with the predicted values at 50 degrees C (K(m) 0.45 mg mL(-1) and V(max) 111.11 micromol mg(-1) protein min(-1)) and at 60 degrees C (K(m) 0.36 mg mL(-1)and V(max) 142.85 micromol mg(-1) protein min(-1)). An Arrhenius plot indicated thermal activation up to 60 degrees C, and thereafter, inactivation. The enzyme was strongly stimulated by Co(2+), Fe(2+), Ag(2+), and Ca(2+), slightly stimulated by Cu(2+) and Mg(2+), and inhibited by Hg(2+), Zn(2+), Ni(2+), and Mn(2+). Among additives, dextran and trehalose slightly enhanced the activity. Glucoamylase activity was inhibited by EDTA, beta-mercaptoethanol, dithiothreitol, and n-bromosuccinimide, and n-ethylmaleimide inhibited its activity completely. This suggested the involvement of tryptophan and cysteine in catalytic activity and the critical role of disulfide linkages in maintaining the conformation of the enzyme. The enzyme hydrolyzed around 82% of soluble starch and 65% of raw starch (K(m) 2.4 mg mL(-1), V(max) 50 micromol mg(-1) protein min(-1)), and it was remarkably insensitive to glucose, suggesting its applicability in starch saccharification.     

Optimization of culture variables for improving glucoamylase production by alginate-entrapped Thermomucor indicae-seudaticae using statistical methods

Alginate-entrapped sporangiospores of Thermomucor indicae-seudaticae were used for the production of glucoamylase. The critical variables that affected glucoamylase production were identified by Plackett-Burman design (sucrose, yeast-extract, K(2)HPO(4) and asparagine) and further optimized by using a four factor central composite design (CCD) of response surface methodology (RSM). Immobilized sporangiospores secreted 41% and 60% higher glucoamylase titers in shake flasks and airlift fermenter, respectively, when the variables were used at their optimum levels (sucrose 3.0%, yeast-extract 0.2%, K(2)HPO(4) 0.1% and asparagine 0.35%). Glucoamylase production (26.3 U ml(-1)) in the optimized medium was in good agreement with the values predicted by the quadratic model (26.7 U ml(-1)), thereby confirming its validity. The enzyme production was sustainable in flasks of higher volume and also airlift fermenter, and attained a peak within 32 h in the fermenter as compared to that of 48 h in shake flasks.     

Bioconversion and binding of sterols by thermophilic moulds

None of the fourteen thermophilic moulds was able to break down the aliphatic side chain of sterols,viz. cholesterol, lanosterol, sitosterol, and stigmasterol so as to yield 4-androstene-3, 17-dione, 1,4-androstadiene-3, 17-dione and progesterone. InAcremonium alabamensis and.Talaromyces emersonii, cholestenone was detected as a product of fermentation of cholesterol whereas the former yielded stigmastadienone from stigmasterol and sitosterol. Lanosterol appeared to be resistant to fungal bioconversion. All the thermophilic moulds exhibited avidity for binding sterols to the mycelium, but the ability to bind sterol seemed to depend upon the nature of the organism and the sterol.     

Lipid composition of cyanidium

The major lipids in Cyanidium caldarium Geitler are monogalactosyl diglyceride, digalactosyl diglyceride, plant sulfolipid, lecithin, phosphatidyl glycerol, phosphatidyl inositol, and phosphatidyl ethanolamine. Fatty acid composition varies appreciably among the lipids, but the major ones are palmitic acid, oleic acid, linoleic acid, and moderate amounts of stearic acid. Trace amounts of other acids in the C₁₄ to C₂₀ range were also present. Moderate amounts of linolenic acid were found in two strains, but not in a third. The proportion of saturated acid is relatively high in all lipids ranging from about a third in monogalactosyl diglyceride to three-fourths in sulfolipid. This may be a result of the high growth temperature. Lipases forming lysosulfolipid, and lysophosphatidyl glycerol are active in ruptured cells; galactolipid is degraded with loss of both acyl residues. Thus the lipid and fatty acid composition of Cyanidium more closely resembles that of green algae than that of the blue-green algae, although there are differences of possible phylogenetic interest.     

Creation of a polyenzyme composition of proteases from Streptomyces griseus and Acremonium chrysogenum

Proteolytic enzymes of microorganisms have been studied for the possibility to create their polyenzymic composition in order to rise a degree of protein hydrolysis and to lower the process duration. Optimal action conditions are selected and a hydrolysis of a number of globular and fibrillar proteins is conducted by a polyenzymic system of Streptomyces griseus and Acremonium chrysogenum proteases.     

Lipid composition of sweet cherries

Lipid comprises about 0.1 % of the fresh weight of cherries. The major components of the neutral lipid fraction are wax and sterol esters, sitosterol, oleanolic and ursolic acids. The glycolipid fraction is comprised of monoglycosyldiacylglycerol, diglycosyldiacylglycerol and acylated sterol glycoside, while phosphatidylcholine and phosphatidylethanolamine are the major phospholipids. Palmitic, oleic, linoleic and linolenic acids comprise over 85% of the total fatty acids in each lipid fraction.     

Lipid composition of housefly microsomes

The lipid composition of microsomes isolated from whole-body preparations of a diazinon-resistant strain of the housefly (Musca domestica L.) was determined. When calculated on a mg% basis housefly microsomes are composed of 47% neutral lipids and 53% phospholipids. The free fatty acids compose the major group of compounds among neutral lipids (60%) and the major phospholipid is phosphatidylethanolamine (47.6%). The molar cholesterol/phospholipid ratio is 0.14, ca. twofold higher than the ratio found in mammalian liver microsomes. Fatty acid composition of housefly microsomes is notable only in that there is no uniform distribution of fatty acid moieties throughout the various classes of lipids, except for linoleic acid (18:2), which accounted for 9.2-12.6% of the neutral lipids. Eighty percent of the fatty acid moieties of phosphatidylserine (+ phosphatidylinositol) were unsaturated. The highest percentage of saturated fatty acid moieties (64.26%) was found in lysophosphatidylethanolamine. Palmitoleic acid (16:1), the major fatty acid found in higher dipterans, ranged from 13.51% in lysophosphatidylethanolamine to 37.45% in the free fatty acids. One prostaglandin (PGF_1d) and leukotriene B₄ were detected in the microsomal lipids at concentrations of 59.7 and 716 pg/200 mg of protein, respectively.     

Lipid composition of Mycoplasma neurolyticum

The total lipid content of Mycoplasma neurolyticum comprises about 14% of the dry weight of the organisms and is about equally distributed between the phospholipid and the neutral-glycolipid fractions. The neutral lipids were identified as triglycerides, diglycerides, and cholesterol. The glycolipid fraction contained 1-O-beta-glucopyranosyl-d-2,3-diglyceride and 1-[O-beta-d-glycopyranosyl-(1-->6)-O-beta-d-glucopyranosyl]-d-2,3-diglyceride. The latter lipid is structurally identical to the diglucosyl diglyceride which occurs in Staphylococcus aureus. The phospholipids of the organism consist of a fully acylated glycerophosphoryl-glycerophosphoryl glycerol, phosphatidic acid, diphosphatidyl glycerol, phosphatidyl glycerol, and amino acyl esters of phosphatidyl glycerol. Phosphatidic acid and phosphatidyl glycerol account for greater than 90% of the phospholipids of organisms in the exponential phase of growth. The predominant fatty acids found in all of the acyl lipids were palmitic, stearic, and oleic acids.     

Lipid composition of the yeast

Summary The lipids of two strains of Lipomyces kononenkoae, grown in batch culture, were extracted and analysed. The major lipids present were phospholipids, free sterols, esterified sterols, and triacylglycerols. Phospholipid analysis indicated that phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol were the major ones. The fatty-acyl residues were C₁₂-C₁₈ and contained 67–74% unsaturated residues. Polyunsaturated residues accounted for 15% and 30% in L. kononenkoae CBS 2514 and L. kononenkoae CBS 5608, respectively. Analysis of the fatty-acyl residues of a low-density vesicle fraction obtained from sphaeroplasts of L. kononenkoae CBS 2514 was carried out and the results are discussed in relation to plasma membrane synthesis. The suitability of L. kononenkoae for production of single-cell protein is also discussed.     

Lipid composition of Halobacterium lacusprofundi

Abstract The recently described aerobic, extremely halophilic archaeobaterium, Halobacterium lacusprofundi was subjected to lipid analysis so that comparisons could be made between existing lipid data and that of the new isolate. This investigation showed that the major respiratory lipoquinones present were MK-8 and MK-8(VIII-H₂), a feature found in other members of the family Halobacteriaceae. The polar lipids comprised the diether derivatives of phosphatidly glycerol, phosphatidyl glycerophosphate, phosphatidly glycerosulphate, a diglycosyl diether and its sulphate derivative. The data presented shows that Halobacterium locusprofundi is related to Hb. saccharovorum and Hb. sodomense , and is in agreement with phylogenetic data.     

The floristic composition of moulds growing on walls of food and drink processing factories

Samples were taken from heavily moulded walls at a range of food and drink processing plants. Quantitative estimates of the number of moulds, yeasts and bacteria were made and the moulds were identified. The floristic diversity of the germination areas and kilns of several maltings establishments was also investigated. The most commonly isolated organisms were Penicillium spp. although each environment appears to have its own characteristic flora with more than 50% of the total colony forming units isolated at each site being accounted for by three or fewer species. The results are discussed in relation to the microflora of mouldy walls in other environments.Several of the fungi associated with the maltings establishments were not found at any other sites. The origin of these fungi is discussed in relation to the flora associated with grain and processing of malt.     

Lipid composition of Trypanosoma cruzi.

1. Lipid composition of Trypanosoma cruzi epimastigote form in culture consist of 35% of phospholipids and 65% of neutral lipids. 2. Among the phospholipids, phosphatidylcholine is the more abundant (44%), followed by phosphatidylethanolamine (28%), phosphatidylinositol (12%), sphingomyelin (4%), and smaller amounts of cardiolipin, phosphatidic acid, lysolecithin, phosphatidylserine (traces), and an unidentified phospholipid (3%). 3. Pulse labeling with 32P showed highest specific incorporation in phosphatidylethanolamine, followed by phosphatidylinositol and phosphatidylcholine, suggesting a more active role for phosphatidylethanolamine in these organisms.