共查询到20条相似文献,搜索用时 15 毫秒
1.
C.P. Kubicek 《FEMS microbiology letters》1983,20(2):285-288
Abstract The effect of l -sorbose on growth, morphology, cell wall composition and β-glucosidase location has been examined with Trichoderma pseudokoningii . Sorbose-grown cultures exhibited a longer lag phase, a tendency to more frequent hyphal branching and showed a decreased cell wall content of β-1,3-glucan. In sorbose-containing cultures, a significant higher portion of total β-glucosidase was present in the culture fluid, whereas in sorbose-lacking control cultures the major part of activity was associated with the cell walls. The results support the previous hypothesis (Kubicek, C.P. (1982) Arch. Microbiol. 132, 349–354) that β-1.3-glucan is involved in cell wall binding of β-glucosidase in Trichoderma pseudokoningii . 相似文献
2.
The localization of β -glucosidase was determined at the tissue level in roots and shoots of rye, wheat and maize seedlings, using an immunohistochemical approach with antibodies directed against purified maize β -glucosidase as the primary antibody. In the roots, the β -glucosidase was found in the epidermis and the underlying cell layer. In the leaves, staining was seen in the epidermis (rye and wheat) and nearby vascular tissue (rye, wheat and maize). In all 3 species, β -glucosidase activity was highest in the coleoptile. Here the enzyme was restricted to the epidermis in wheat and to cells near the vascular tissue in maize, but was found in the whole tissue, except the vascular tissue, in rye. Maize, wheat and rye all contain hydroxamic acid glucosides and results are discussed in relation to a proposed role of β -glucosidase as part of a defense system releasing hydroxamic acid aglucone upon herbivore attack, pathogen penetration or aphid infestation. 相似文献
3.
Ken Horii Takashi Adachi Takanori Tanino Tsutomu Tanaka Hiroshi Sahara Seiji Shibasaki Chiaki Ogino Yoji Hata Mitsuyoshi Ueda Akihiko Kondo 《Biotechnology letters》2009,31(8):1259-1264
A molecular display technology that uses the displayed proteins on cell surfaces has many applications in microbiology and
molecular biology. Here, we describe the resistance of displayed proteins to proteases using simulated gastric fluid (SGF),
which included pepsin at pH 2. The displayed β-glucosidase resisted pepsin digestion compared with secreted, free β-glucosidase.
In SDS-PAGE and Western blotting analysis, the secreted β-glucosidase was immediately digested within 1 min following SGF
treatment, although the displayed β-glucosidase was stable for more than 60 min following SGF treatment. In addition, the
residual activity of secreted β-glucosidase was completely destroyed after 10 min SGF treatment. However, displayed β-glucosidase
retained 14% of its residual activity following the same treatment. These results clearly show that cell surface display technology
using enzymes can reveal the protease resistance of a protein of interest under various conditions. 相似文献
4.
Abstract A Monilia sp. produced an inducible intracellular β- d -glucosidase (IG-2) which is the nascent form of the extracellular enzyme (EG-1) prior to its secretion into the extracellular medium. The other intracellular β- d -glucosidase (IG-1) produced was a constitutive enzyme. Highest yields of the inducible β- d -glucosidase resulted when Monilia sp. was grown on insoluble cellulose. Cellobiose and d -glucose appeared to repress β- d -glucosidase formation at high substrate levels and synthesis occurred only once the levels of these sugars in the medium were nearly depleted. 相似文献
5.
Purification and characterization of an intracellular β-glucosidase from a new strain of Leuconostoc mesenteroides isolated from cassava 总被引:1,自引:1,他引:0
Y. Gueguen P. Chemardin P. Labrot A. Arnaud & P. Galzy 《Journal of applied microbiology》1997,82(4):469-476
The lactic acid bacterium, Leuconostoc mesenteroides, when grown on an arbutin-containing medium, was found to produce an intracellular β-glucosidase. The enzyme was purified by chromatofocusing, ion-exchange chromatography and gel filtration. The molecular mass of the purified intracellular β-glucosidase, as estimated by gel filtration, was 360 kDa. The tetrameric structure of the β-glucosidase was determined following treatment of the purified enzyme with dodecyl sulphate (SDS). The intracellular β-glucosidase exhibited optimum catalytic activity at 50°C and pH 6 with citrate–phosphate buffer, and 5·5 with phosphate buffer. The enzyme was active against glycosides with (1→4)-β, (1→4)-α and (1→6)-α linkage configuration. From Lineweaver–Burk plots, K m values of 0·07 mmol l−1 and 3·7 mmol l−1 were found for p -nitrophenyl-β- D -glucopyranoside and linamarin, respectively. The β-glucosidase was competitively inhibited by glucose and by D -gluconic acid–lactone and a glucosyl transferase activity was observed in the presence of ethanol. The β-glucosidase of Leuconostoc mesenteroides, with cyanogenic activity, could be of potential interest in cassava detoxification, by hydrolysing the cyanogenic glucosides present in cassava pulp. 相似文献
6.
Few bacteria are capable of degrading crystalline cellulose but there is considerable interest in the properties of enzyme systems with this capability. In the bovine and ovine rumen the principal cellulolytic bacterium is Fibrobacter (formerly Bacteroides ) succinogenes. The cellulase system of this organism is composed of multiple enzyme components, including a constitutive and cell-associated β -glucosidase active against cellobiose. The properties of the β -glucosidase activity have been investigated with the chromogenic substrate β -nitrophenyl β -D-glucoside (pNPG). Hydrolytic activity against pNPG was located primarily in the cytoplasm and the cytoplasmic membrane but showed a gradual migration to the periplasm during growth on either glucose or cellobiose. Activity against cellobiose was found in the periplasm in significant amounts in all growth phases. Of the β -glucosides tested, only cellobiose and pNPG were hydrolysed by crude cell extracts. In the presence of cellobiose, however, the rate of hydrolysis of pNPG was stimulated up to 10-fold, and extracts hydrolysed methylumbelliferyl β -D-glucoside, 5-bromo-4-chloro-3-indolyl β -D-glucoside, arbutin and aesculin. Activities against pNPG in the presence and absence of cellobiose displayed similar instability in the presence of oxygen; both were stabilized by dithiothreitol and the temperature and pH optima were identical. A significant proportion of the membrane-associated β -glucosidase was released by treatment with 0.3 mol/1 KCl, and fractionation by chromatography on CM-cellulose showed the presence of two activities against pNPG, only one of which was stimulated by cellobiose. 相似文献
7.
Christel Béra Véronique Broussolle Evelyne Forano Geneviève Gaudet 《FEMS microbiology letters》1996,136(1):79-84
Abstract β-Glucosidase in Aspergillus nídulans was found to be both intracellular and extracellular. The intracellular β-glucosidase was synthesized after the exhaustion of carbon source in the medium. The extracellular enzyme appeared with autolysis of the mycelium. Biosynthesis of β-glucosidase was not induced by various carbohydrates but repressed to varying extents in the presence of glucose, glycerol, and 2-deoxyglucose. This repression was not relieved by addition of cAMP. The repression was relieved much more by mutations in the creA gene than by one in the creC gene. Thus, β-glucosidase synthesis in A. nidulans is subject to carbon catabolite repression. 相似文献
8.
GLYCOSIDASES IN NORMAL AND SCRAPIE MOUSE BRAIN 总被引:4,自引:2,他引:2
Abstract— The pH optima of ten glycosidases have been determined in normal and scrapie-affected mouse brain. The enzymes α-mannosidase, α-glucosidase and β-glucosidase displayed two peaks of enzyme activity over the pH range examined.
There is a significant increase in the activity of the enzymes α-mannosidase, β-glucuronidase, N -acetyl β-D-glucosaminidase, N -acetyl β-galactosaminidase, β-glucosidase (pH 4.1), α-fucosidase and β-xylosidase in the brains of mice clinically affected with scrapie, whilst only α-mannosidase (pH 4.1), β-glucuronidase, N -acetyl-β-D-glucosinidase and N- acetyl-β-D-gaiactosaminidase are elevated before mice exhibit signs of the disease. 相似文献
There is a significant increase in the activity of the enzymes α-mannosidase, β-glucuronidase, N -acetyl β-D-glucosaminidase, N -acetyl β-galactosaminidase, β-glucosidase (pH 4.1), α-fucosidase and β-xylosidase in the brains of mice clinically affected with scrapie, whilst only α-mannosidase (pH 4.1), β-glucuronidase, N -acetyl-β-D-glucosinidase and N- acetyl-β-D-gaiactosaminidase are elevated before mice exhibit signs of the disease. 相似文献
9.
Abstract The cellulolytic actinomycete Streptomyces sp. QM-B814 posasses an intracellular β-glucosidase system which is induced by cellobiose and carboxymethylcellulose. Maximal β-glucosidase activity was attained 8–10 h after inducer addition to exponential phase growing cultures. The induction is depressed in the presence of glucose. The system is composed of two electrophoretically different β-glucosidase forms showing relative molecular masses of about 60 and 35 kDa, and p I values in the range 4.2–4.5. Both β-glucosidases are synthesized de novo. The enzymes share substrate preference and are both inhibited by δ-gluconolactone and p -chloromercuribenzoate. The induction pattern and glucose inhibition are similar for both enzymes. 相似文献
10.
The cDNA for a β-glucosidase (EC3.2.1.21) was isolated from rye ( Secale cereale , cv Motto) and the sequence corresponding to the mature protein cloned into pET21a expression vector and used for transformation of Escherichia coli. The recombinant β-glucosidase expressed in E. coli was recognized by antibodies to maize β-glucosidase and exhibited the same kinetic properties on the endogenous substrates hydroxamic acid glucosides and artificial substrates as the native enzyme purified from rye. The enzyme monomer had an apparent molecular weight of about 67 kDa. The isolated cDNA was analysed with web-based chloroplast targeting prediction programs. The programs predicted a chloroplast targeting peptide with a cleavage site between amino acid 49 and 50. Sequence alignment of the plastidic rye β-glucosidase showed that the putative sites for substrate specificity of maize Glu1, W378 and F198 (F197) are conserved in the rye enzyme, whereas F205, F466 and A467 of maize Glu1 are exchanged for histidine, glycine and serine, respectively, in rye. The plastidic β-glucosidase is expressed in all plant parts and the highest levels were found in the coleoptile and mesocotyl. 相似文献
11.
Use of perfused cores for evaluating extracellular enzyme activity in stream-bed sediments 总被引:2,自引:0,他引:2
Abstract β-Glucosidase activity was investigated in stream-bed sediments using 4-methylumbelliferyl-β- d -glucopyranoside (MUF-β-Glc) as a model substrate. In a perfused core technique, water containing MUF-β-Glc was perfused up through sediment cores. β-glucosidase activity quantified from the release of fluorescent MUF in water discharge from the cores. At low rates of perfusion, maximum β-glucosidase activity ( V max ) in perfused sediments was similar to that in suspended (unperfused) sediments. Substrate affinity( K m)was higher in the suspended sediments. V max and K m both increased when the perfusion rate was raised, although naturally-low substrate concentrations could mean that variability in perfusion rates has little effect on enzyme activity in the field. V max was uninfluenced by whether ground or stream water was perfused through the sediments, but K m was higher in cores perfused with groundwater. Increasing concentrations of glucose in the perfusion water resulted in a progressive inhibition of β-glucosidase activity. Although natural concentrations of glucose were low, the high turnover of enzymatically-released glucose probably means that β-glucosidase activity could be regulated by product concentration. 相似文献
12.
13.
Hydrolytic enzyme(s) production in Micrococcus roseus growing on different cellulosic substrates 总被引:1,自引:0,他引:1
Micrococcus roseus (G12) isolated from higher termite Odontotermes obesus gut exhibited cellulose digesting properties. A lignocellulosic substrate, rice husk induced endoglucanase, β-glucosidase, β-xylanase and β-xylosidase production. Besides rice husk, CMC also induced endoglucanase production. β-Glucosidase activity was quite pronounced when rice husk was supplemented with CMC or cellobiose. Both β-xylanase and β-xylosidase activities could be induced by xylan as well as xylobiose, whereas CMC induced partial activity. Endoglucanase and β-xylanase enzymes were secreted into the culture medium, whereas β-glucosidase and β-xylosidase activities were intracellular in nature. Enzyme production was subject to end product inhibition. The extracellular enzyme(s) possessed the potential to saccharify rice husk, xylan and CMC to reducing sugars. 相似文献
14.
Variations in faecal bacterial enzyme activities and associations with bowel function and diet in elderly subjects 总被引:1,自引:0,他引:1
Daily and inter-individual variations of faecal bacterial β-glucuronidase and β-glucosidase activities and their associations with parameters of bowel function were studied in 10 residents of an old people's home during two 1-week periods 2 weeks apart. The effect of sampling method (a spot sample vs an aliquot of the homogenized sample from a total daily collection) on the activities of these enzymes and that of urease was also assessed. Intestinal transit time was determined using the radio-opaque Sitzmark® ; capsules, and questionnaires on bowel function and intakes of fluids and fibre-containing foods were completed. The mean (95% confidence interval) β-glucuronidase and β-glucosidase levels were 3·08 (2·75–3·41) and 11·53 (10·79–12·26) nmol min−1 mg protein−1 . Daily variations in enzyme activities within individuals were not significant ( P = 0·277 and 0·990, respectively), whilst those between individuals were highly significant ( P = 0·000). Faecal frequency correlated negatively with β-glucuronidase and urease, but no other associations of the enzymic activities with parameters of bowel function and diet were observed. β-Glucuronidase and β-glucosidase were not affected by the sampling method, while significantly higher urease was obtained by spot sampling as compared with the aliquot representing the total daily collection. Large inter-individual variations in faecal enzyme activities should be taken into consideration when planning experiments and interpreting results on these faecal parameters. 相似文献
15.
The influence of different cultivation conditions on β-glucosidase production and of some parameters on the activity and stability
of this enzyme were studied inNectria catalinensis. Maximal β-glucosidase production was achieved with ammonium nitrate (0.5 g N/L) as nitrogen source. Tween 80, Tween 20 and
Triton X-100 increased β-glucosidase yields, Tween 80 was the most effective for enzyme release and growth at a concentration
of 3.4 mmol/L. On the other hand, Tween 20 and Triton X-100 had an inhibitory effect onN. catalinensis growth. A temperature of 23°C and an initial pH of cultures of 6.5 were optimal for biomass and β-glucosidase production.
Under optimal cultural conditions (ammonium nitrate, 0.5 g N/L; Tween 80, 3.4 mmol/L; 23°C; initial pH 6.5) the β-glucosidase
yield was increased more than five fold respect to the initial state. Optimal temperature for β-glucosidase activity was 45°C,
the initial activity dropped 60 % after 6 h of incubation at this temperature. Optimal pH for enzyme activity was 5.3. At
this pH the β-glucosidase was completely stable after 3 d of incubation. TheV andK
m values calculated from Lineweaver-Burk and Eadie-Hofstee plots were 0.23 μmol 4-nitrophenol per min per mg of protein and
0.25 mmol 4-nitrophenol β-d-glucopyranoside per L, respectively. The activation energy according to Arrhenius plot was 49.6 KJ/mol. 相似文献
16.
The subcellular compartmentation of β -glucosidase was studied in rye, maize and wheat seedlings by immunocytochemical methods. For detection, we used a 10 nm gold-labeled secondary antibody, and results were observed using transmission electron microscopy. In all three species, β -glucosidase was found in plastids, cytoplasm and cell walls. In rye, gold particles were seen on cell walls and cytoplasm in epidermal cells of the root tip and shoot, in bundle sheath cells of the shoot and in all cells, except the vascular bundle cells of the coleoptile. Gold labeling was also observed in plastids of the bundle sheath cells of rye shoot tips and in cortical cells of root tips. In wheat, gold labeling was observed on cell walls and cytoplasm of epidermal cells in the shoot base and coleoptile, and on cell walls and plastids in epidermal cells of the root tip. In maize, gold labeling was mainly found in plastids or proplastids in vascular bundle cells and bundle sheath cells of the shoot, in bundle sheath cells of the coleoptile and in epidermal cells of the root. Some gold particles were also found in cell walls and cytoplasm of stomatal guard cells of the shoot base and vascular bundle cells of the shoot tip and in the cell walls of bundle sheath cells of the shoot tip and root tip epidermal cells. Results are discussed in relation to the role of β -glucosidase in hydroxamic acid release and overall defense mechanism of monocotyledons. 相似文献
17.
Abstract Genomic DNA fragments encoding β-glucosidase activity from the wild-type strain WD4 of Erwinia herbicola were cloned into Escherichia coli . Two clones containing a common fragment encoded a polypeptide of 58000 Da. Cloned β-glucosidase, expressed in E. coli , showed activity against natural β-glucoside sugars except for cellobiose. An open reading frame of 1442 bp termed bglA was identified by nucleotide sequencing and it coded for a protein of 480 amino acids ( M r 53896) which showed significant homology with β-glucosidases from glycosyl hydrolase family 1. 相似文献
18.
A β-glucosidase gene (bglI) from Trichoderma reesei was cloned into the pPIC9 vector and integrated into the genome of Pichia pastoris GS115. Under the control of the methanol-inducible alcohol oxidase (AOX) promoter and using Saccharomyces
cerevisiae secretory signal peptide (α-factor), the recombinant β-glucosidase was expressed and secreted into the culture medium. The
maximum recombinant β-glucosidase activity achieved was 60 U/ml, and β-glucosidase expression reached 0.3 mg/ml. The recombinant
76 kDa β-glucosidase was purified 1.8-fold with 26% yield and a specific activity of 197 U/mg. It was optimally active at
70°C and pH 5.0. 相似文献
19.
Hong SY Lee JS Cho KM Math RK Kim YH Hong SJ Cho YU Cho SJ Kim H Yun HD 《Biotechnology letters》2007,29(6):931-936
An artificial bifunctional enzyme, cellulase-β-glucosidase, was prepared by gene fusion from the hyperthermophilic bacterium
Thermotoga maritima MSB8. The fusion protein exhibited both cellulase (Cel5C) and β-glucosidase (BglB) activity when the bglB gene was fused to downstream of cel5C, but not when cel5C was fused to downstream of bglB. The specific activity of the bifunctional enzyme was 70% lower than that of cellulase or β-glucosidase. The fusion enzyme
was purified, and the MW was estimated as 114 kDa. The fusion enzyme displayed optimum cellulase activity at pH 8.0 and 70°C
over 30 min, and optimal β-glucosidase activity at pH 7.0 and 80°C over 30 min. 相似文献
20.
Xylanase, β-glucosidase, β-xylosidase, endoglucanase and polygalacturonase production fromCurvularia inaequalis was carried out by means of solid-state and submerged fermentation using different carbon sources. β-Glucosidase. β-xylosidase,
polygalacturonase and xylanase produced by the microorganisms were characterized. β-Glucosidase presented optimum activity
at pH 5.5 whereas xylanase, poly-galacturonase and β-xylosidase activities were optimal at pH 5.0. Maximal activity of β-glucosidase
was determined at 60°C, β-xylosidase at 70°C, and polygalacturonase and xylanase at 55°C. These enzymes were stable at acidic
to neutral pH and at 40–45 °C. The crude enzyme solution was studied for the hydrolysis of agricultural residues. 相似文献