共查询到20条相似文献,搜索用时 46 毫秒
1.
Yu Qiu Hai Niu Wen Huang Yang He Xiao-Hua Wu 《Biotechnology and Bioprocess Engineering》2011,16(5):858-866
A tannase with a molecular mass of 72 kDa was obtained from Penicillium herquei isolated from valonia acorns following fermentation in a 5 L bioreactor. This tannase showed optimum activity at pH 6.0 and
30°C. The enzyme was inhibited by Fe3+, Zn2+, dithiothrietol (DTT), β-mercaptoethanol, formaldehyde, and ethanol, and induced by K+, Mn2+, Tween 80, and Triton X-100. The Michaelis constant (K
m) and the second-order constant (k
cat/K
m) values of the tannase for propyl gallate (PG) were 0.62 mM and 174.1 mM/sec. The circular dichroism (CD) spectra indicated
that the secondary structure of the tannase contained 14% α helix, 32.4% anti-parallel β-sheet, 4.8% β-sheet, 18.8% β-turn,
and 30% random coil. Native tannase in ultrapure water manifested as spherical nano-particle aggregates with diameters ranging
from 50 to 300 nm determined by atomic force microscopy (AFM). 相似文献
2.
A thermoalkaliphilic lipase of <Emphasis Type="Italic">Geobacillus</Emphasis> sp. T1 总被引:3,自引:0,他引:3
Leow TC Rahman RN Basri M Salleh AB 《Extremophiles : life under extreme conditions》2007,11(3):527-535
A thermoalkaliphilic T1 lipase gene of Geobacillus sp. strain T1 was overexpressed in pGEX vector in the prokaryotic system. Removal of the signal peptide improved protein
solubility and promoted the binding of GST moiety to the glutathione-Sepharose column. High-yield purification of T1 lipase
was achieved through two-step affinity chromatography with a final specific activity and yield of 958.2 U/mg and 51.5%, respectively.
The molecular mass of T1 lipase was determined to be approximately 43 kDa by gel filtration chromatography. T1 lipase had
an optimum temperature and pH of 70°C and pH 9, respectively. It was stable up to 65°C with a half-life of 5 h 15 min at pH
9. It was stable in the presence of 1 mM metal ions Na+, Ca2+, Mn2+, K+ and Mg2+ , but inhibited by Cu2+, Fe3+ and Zn2+. Tween 80 significantly enhanced T1 lipase activity. T1 lipase was active towards medium to long chain triacylglycerols (C10–C14)
and various natural oils with a marked preference for trilaurin (C12) (triacylglycerol) and sunflower oil (natural oil). Serine
and aspartate residues were involved in catalysis, as its activity was strongly inhibited by 5 mM PMSF and 1 mM Pepstatin.
The T
m for T1 lipase was around 72.2°C, as revealed by denatured protein analysis of CD spectra. 相似文献
3.
Yoshiyuki Takasaki 《Bioscience, biotechnology, and biochemistry》2013,77(6):1539-1547
An α-amylase which produces maltohexaose as the main product from strach was found in the culture filtrate of Bacillus circulans G-6 which was isolated from soil and identified by the author.The enzyme was purified by means of ammonium sulfate fractionation, DEAE-Sepharose column chromatography and Sephadex G-200 column chromatography. The purified enzyme was homogeneous on disc electrophoresis. The optimum pH and temperature of the enzyme were around pH 8.0 and around 60°C, respectively. The enzyme was stable in the range of pH 5–10. Metal ions such as Hg2+, Cu2+, Zn2+, Fe2+ and Co2+ inhibited the enzyme activity. The molecular weight was about 76,000. The yield of maltohexaose from soluble starch of DE (dextrose equivalent*) 1.8-12.6 was about 30%, and the combined action of the enzyme and pullulanase or isoamylase increased the yield of maltohexaose. 相似文献
4.
A newly discovered bacterium, strain AC1, containing cellulase was isolated from the gastric juice of the mollusca, Ampullaria crosseans. Analysis of the 16S rDNA sequence and carbon sources revealed that the bacterium belonged to the genus Bacillus. A novel endoglucanase (Ba-EGA) was purified from culture supernatants of the bacterium growing in CMC-Na (low viscosity) induction medium. The cellulase
was purified about 150-fold by ammonium sulfate fractionation, ion exchange, hydrophobic, and gel filtration chromatography,
with a specific activity of 35.0 IU/mg. The molecular mass of the enzyme was 67 kDa. N-terminal amino acid sequencing revealed
a sequence of SDYNYVEVLQKSILF, which had high homology with endoglucanases from the Bacillus and Clostridium species. The maximal activity of the enzyme with the substrate of CM-cellulose is at pH 4.5–6.5 and 70°C, respectively. The
studies on pH and temperature stability showed that the Ba-EGA is stable enough between pH 7.5 and 10.5 at 30°C for 2 h, and more than 80% of the activity still remains when incubation
was prolonged to 1 h at 50°C. The activity of the enzyme was significantly inhibited by Fe2+, Cu2+ (5.0 mM of each), and sodium dodecyl sulfate (SDS) (0.5%) and obviously activated by Tween 20 and Triton X-100 (0.25% each).
Binding studies revealed that the Ba-EGA had cellulose-binding domain. 相似文献
5.
Streptococcus dysgalactiae IID 678, belonging to group C of the streptococci, secreted a large amount of hyaluronidase (hyaluronate lyase, EC 4.2.2.1) into a culture medium containing hyaluronic acid. The purification procedures of hyaluronidase were 70% ammonium sulfate precipitation, ECTEOLA-cellulose chromatography, phospho-cellulose chromatography, and gel filtration on Sephacryl S-300. The hyaluronidase was purified approximately 27,000-fold from the culture filtrate. The purified enzyme was homogeneous by SDS-poIyacrylamide gel electrophoresis. The enzyme degradated only hyaluronic acid and chondroitin to zl 4,5-unsaturated disaccharides and did not act on other glycosaminoglycans containing sulfate groups, while the degradation rate of chondroitin was about 1/60 of that of hyaluronic acid. The optimum pH was wide, from pH 5.8 to pH 6.6, and the optimum temperature was 37°C. Fe2 +, Cu2 +, Pb2 +, and Hg2 + ions inhibited the activity strongly and Zn2+ inhibited it by half. The molecular weight of the enzyme was estimated to be 125,000 by gel filtration and 117,000 by SDS-polyacrylamide gel electrophoresis. The enzyme was different immunochemically from the hyaluronidase from Streptococcus pyogenes belonging to group A. 相似文献
6.
Shinji Yoshitake Kei-ichi Watanabe Gunki Funatsu 《Bioscience, biotechnology, and biochemistry》2013,77(10):2193-2195
An α-amylase which produces both maltotetraose and maltopentaose from starch as the main products was found in the culture filtrate of a strain of Bacillus circulans which was newly isolated from soil. The enzyme was purified to be almost homogeneous on disc electrophoresis in polyacrylamide gel by means of ammonium sulfate fractionation, DEAE-Sepharose column chromatography and Sephadex G-200 gel filtration.The optimum pH and temperature of the enzyme were around pH 7.0 and around 50°C, respectively. Metal ions such as Hg2+, Cu2+, Ni2+, Zn2+, Fe2+ and Co2+ strongly inhibited the enzyme activity. The molecular weight was about 45,000. The yields of maltotetraose and maltopentaose from potato starch were 30 ~ 40% and 20 ~ 30%, respectively. 相似文献
7.
Hana Maalej Noomen Hmidet Olfa Ghorbel-Bellaaj Moncef Nasri 《Biotechnology and Bioprocess Engineering》2013,18(5):878-887
This study reports the purification and biochemical characterization of a novel maltotetraose-forming-α-amylase from Pseudomonas stutzeri AS22, designated PSA. The P. stutzeri α-amylase (PSA) was purified from the culture supernatant to homogeneity by Sepharose mono Q anion exchange chromatography, ultrafiltration and Sephadex G-100 gel filtration, with a 37.32-fold increase in specific activity, and 31% recovery. PSA showed a molecular weight of approximately 57 kDa by SDS-PAGE. The N-terminal amino acid sequence of the first 7 amino acids was DQAGKSP. This enzyme exhibited maximum activity at pH 8.0 and 55°C, performed stably over a broad range of pH 5.0 ≈ 12.0, but rapidly lost activity above 50°C. Both potato starch and Ca2+ ions have a protective effect on the thermal stability of PSA. The enzyme activity was inhibited by Hg2+, Mn2+, Cd2+, Cu2+, and Co2+, and enhanced by Ba2+. PSA belonged to the EDTA-sensitive α-amylase. The purified enzyme showed high stability towards surfactants (Tween 20, Tween 80 and Triton X-100), and oxidizing agents, such as sodium per borate and H2O2. In addition, PSA showed excellent compatibility with a wide range of commercial solid and liquid detergents at 30°C, suggesting potential application in the detergent industry. Maltotetraose was the specific end product obtained after hydrolysis of starch by the enzyme for an extended period of time, and was not further degraded. 相似文献
8.
Sung-Il Kang Young-Boo Jang Yeung-Joon Choi Jai-Yul Kong 《Biotechnology and Bioprocess Engineering》2005,10(6):593-598
A collagenolytic enzyme, produced byVibrio vulnificus CYK279H, was purified by ultrafiltration, dialysis, Q-Sepharose ion exchange and Superdex-200 gel chromatography. The enzyme
from the supernatant was purified 13.2 fold, with a yield of 11.4%. The molecular weight of the purified enzyme was estimated
by SDS-PAGE to be approximately 35.0 kDa. The N-terminal sequence of the enzyme was determined as Gly-Asp-Pro-Cys-Met-Pro-Ile-Ile-Asn.
The optimum temperature and pH for the enzyme activity were 35°C and 7.5, respectively. The enzyme activity was stable within
the pH and temperature ranges 6.8∼8.0 and 20∼35°C, respectively. The purified enzyme was strongly activated by Zn2+, Li2+, and Ca2+, but inhibited by Cu2+. In addition, the enzyme was strongly inhibited by 1, 10-phenanthroline and EDTA. The purified enzyme was suggested to be
a neutral metalloprotease. 相似文献
9.
Characterization of a recombinant thermostable xylanase from deep-sea thermophilic Geobacillus sp. MT-1 in East Pacific 总被引:1,自引:0,他引:1
A novel xylanase-producing thermophilic strain MT-1 was isolated from a deep-sea hydrothermal field in east Pacific. A xylanase gene encoding 331 amino-acid peptide from this isolate was cloned and expressed in Escherichia
coli. The recombinant xylanase exhibited maximum activity at 70°C and had an optimum pH of 7.0. It was active up to 90°C and showed activity over a wide pH ranging from 5.5 to 10.0. The crude xylanase presented similar properties in temperature and pH to those of the recombinant xylanase. The recombinant xylanase was stable in 1 mM of enzyme inhibitors (PMSF, EDTA, 2-ME or DTT) and in 0.1% detergents (Tween 20, Chaps or Triton X-100), whereas, it was strongly inhibited by sodium dodecyl sulfate (SDS) (1 mM). In addition, its catalytic function was stable in the presence of Li+, Na+ or K+. However, it was strongly inhibited by Ni2+, Mn2+, Co2+, Cu2+, Zn2+, Cd2+, Hg2+ and Al3+ (1 or 0.1 mM). The K
m and V
max of the recombinant xylanase for oat spelt xylan were calculated to be 1.579 mg/ml and 289 μmol/(min • mg), respectively. Our study, therefore, presented a rapid overexpression and purification of xylanase from deep-sea thermophile aimed at improving the enzyme yield for industrial applications and scientific research. 相似文献
10.
Bahar Yilmaz Mustafa Ozkan Baltaci 《Journal of enzyme inhibition and medicinal chemistry》2016,31(6):1241-1247
In this study, the extracellular thermostable alkaline protease out of A10 strain was purified 1.38-fold with 9.44% efficiency through the ammonium sulfate precipitation-dialysis and DE52 anion exchange chromatography methods. The molecular weight of the enzyme in question along with sodium dodecyl sulfate-polyacrylamide gel electrophoresis was determined to be approximately 40.55?kDa, whereas the optimum pH and temperature ratings were identified as 9.0 and 70?°C, respectively. It was seen that the enzyme had remained stable between pH 7.5–10.5 range, protecting more than 90% of its activity in the wake of 1?h incubation at 60–70?°C. It was also observed that the enzyme enhanced its activity in the presence of Mg2+, Mn2+, K+, while Fe2+, Ni2+, Zn2+, Ag+?and Co2+? decreased the activity. Ca2+, however, did not cause any change in the activity. The enzyme was seen to have been totally inhibited by phenylmethylsulfonyl fluoride, therefore, proved to be a serine alkaline protease. 相似文献
11.
《Journal of Fermentation and Bioengineering》1991,71(4):226-229
An alkalophilic strain isolated from soil produced intracellular cyclomaltodextrinase on the culture medium at an initial pH of 10.6. The strain was identified as closely resembling Bacillus circulans. The enzyme was purified 252-fold from the cell extract by chitosan treatment, ammonium sulfate fractionation, DEAE-Toyopearl column chromatography, and gel filtration. The pH and temperature optima of the purified enzyme were 6.0 and 50°C. The molecular weight of the enzyme was 126,000, with two subunits of 67,000. The isoelectric point was pH 4.2. Enzyme activity was inhibited by Ag+, Hg2+, Cu2+, and p-chloromercuribenzoate. The enzyme hydrolyzed α-, β-, and γ-cyclodextrins, as well as linear maltodextrins, to yield maltooligosaccharides. Starch and maltose were not degraded by the enzyme. 相似文献
12.
Daniel Guerra Franco Regiane Nogueira Spalanzani Emmly Ernesto Lima Clarice Rossato Marchetti Patrícia Oliveira Silva Douglas Chodi Masui 《Biocatalysis and Biotransformation》2017,35(4):249-259
The proteases are enzymes produced by several filamentous fungi with important biotechnological applications. In this work, a protease from Aspergillus flavus was characterized. The culture filtrate of A. flavus was purified to homogeneity by Sephacryl S-200 column chromatography followed by CM–cellulose. The molecular weight of the purified enzyme was estimated to be approximately 32?kDa by SDS–PAGE. The enzyme hydrolysed BTpNA (N-α-benzoyl-dl-tyrosyl-p-nitroanilide), azo-casein and casein as substrates. Optimal temperature and pH were 55?°C and 6.5, respectively. The enzyme was stimulated by Mg2+, Ca2+, Zn2+ and inhibited by Hg2+ and Ag2+ and Cu2+. The protease showed increased activity with detergents, such as Tween 80 and Triton X, and was stable to the reducing agents, such as β-mercaptoethanol. The protease activity was strongly inhibited in the presence of phenylmethylsulfonyl fluoride, indicating it is a serine protease. The enzyme entrapped in calcium alginate beads retained its activity for longer time and could be reused up to 10 times. The thermostability was increased after the immobilization and the enzyme retained 100% of activity at 45?°C after 60?min of incubation, and 90% of residual activity at 50?°C after 30?min. In contrast, the free enzyme only retained 10% of its residual activity after 60?min at 50?°C. The enzymatic preparation was demonstrated to be efficient in the capability of dehairing without destruction of the hide. The remarkable properties such as temperature, pH and immobilization stability found with this enzyme assure that it could be a potential candidate for industrial applications. 相似文献
13.
Małgorzata Pleszczyńska Adrian Wiater Marcin Skowronek Janusz Szczodrak 《Preparative biochemistry & biotechnology》2013,43(4):335-347
Mutanases are enzymes that catalyze hydrolysis of α-1,3-glucosidic bonds in various α-glucans. One of such glucans, mutan, which is synthesized by cariogenic streptococci, is a major virulence factor for induction of dental caries. This means that mutan-degrading enzymes have potential in caries prophylaxis. In this study, we report the purification, characterization, and partial amino acid sequence of extracellular mutanase produced by the MP-1 strain of Paenibacillus curdlanolyticus, bacterium isolated from soil. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme showed a single protein band of molecular mass 134 kD, while native gel filtration chromatography confirmed that the enzyme was a monomer of 142 kD. Mutanase showed a pH optimum in the range from pH 5.5 to 6.5 and a temperature optimum around 40–45°C. It was thermostable up to 45°C, and retained 50% activity after 1 hr at 50°C. The enzyme was fully stable at a pH range of 4 to 10. The enzyme activity was stimulated by the addition of Tween 20, Tween 80, and Ca2+, but it was significantly inhibited by Hg2+, Ag+, and Fe2+, and also by p-chloromercuribenzoate, iodoacetamide, and ethylenediamine tetraacetic acid (EDTA). Mutanase preparation preferentially catalyzed the hydrolysis of various streptococcal mutans and fungal α-1,3-glucans. It also showed binding activity to insoluble α-1,3-glucans. The N-terminal amino acid sequence was NH2-Ala-Gly-Gly-Thr-Asn-Leu-Ala-Leu-Gly-Lys-Asn-Val-Thr-Ala-Ser-Gly-Gln. This sequence indicated an analogy of the enzyme to α-1,3-glucanases from other Paenibacillus and Bacillus species. 相似文献
14.
Ponnambalam Subhashini Neelamegam Annamalai Ayyappan Saravanakumar Thangavel Balasubramanian 《Biologia》2012,67(4):629-635
An extracellular alkaline protease-producing Vibrio sp. was isolated from mangrove sediments of Vellar estuary. A 9.36-fold purification was achieved by a three-step purification procedure and the molecular weight of the enzyme was determined as 33 kDa by SDS-PAGE. The enzyme was active in a broad range of pH (6.0–11.0) and temperature (30–70°C), the optimum being at pH 9.0 and temperature 55°C. The enzyme was stable at alkaline pH range of 9–11 and up to a temperature of 60°C, after incubation for 1 h. Metals like Co2+, Hg2+, Ni2+ and Cu2+ inhibited the enzyme activity, whereas Fe2+, Ca2+ and Mn2+ were found to enhance the activity. The protease was found to be highly stable in the presence of oxidizing agents like H2O2, detergents such as SDS and Triton-X-100 and also some of the commonly used commercial detergents. The organic solvents like xylene, isopropanol, hexane and benzene were found to enhance as well as stabilize the enzyme activity. The extracellular production of the enzyme, the pH and thermal stability, and the stability in presence of oxidants, surfactants, commercial detergents and organic solvents, altogether suggest that it can be used as a laundry additive. 相似文献
15.
Long Liu Hyun‐dong Shin Rachel R. Chen Jianghua Li Guocheng Du Jian Chen 《Biotechnology progress》2013,29(1):39-47
An alkaline α‐amylase gene from alkaliphilic Alkalimonas amylolytica was synthesized based on the preferred codon usage of Escherichia coli and Pichia pastoris, respectively, and then was expressed in the according heterologous host, E. coli BL21 (DE3) and P. pastoris GS115. The alkaline α‐amylase expressed in E. coli was designated AmyA, whereas that produced by P. pastoris was designated AmyB. The specific activity of AmyA and AmyB was 16.0 and 16.6 U/mg at pH 9.5 and 50°C, respectively. The optimal pH and pH stability of AmyA and AmyB were similar, whereas the optimum temperature and thermal stability of AmyB were slightly enhanced compared with those of AmyA. The AmyA and AmyB had a similar melting temperature of 64°C and the same catalytic efficiency (kcat/Km) of 2.0 × 106 L/(mol min). AmyA and AmyB were slightly activated by 1 mM Co2+, Ca2+, or Na+, but inhibited by all other metal ions (K+, Mg2+, Fe3+, Fe2+, Zn2+, Mn2+, and Cu2+). Tween 80 or Tween 60 (10% (w/v)) had little influence on the stability of AmyA and AmyB, while the 10% (w/v) sodium dodecyl sulfate caused the complete loss of AmyA and AmyB activities. The AmyA and AmyB were stable in the presence of solid detergents (washing powder), while were less stable in liquid detergents. Under the optimal conditions in 3‐L bioreactor, the extracellular AmyB activity reached 600 U/mL, which was about 10 times as that of AmyA. These results indicated that P. pastoris was a preferable host for alkaline α‐amylase expression and the produced alkaline α‐amylase had a certain application potential in solid detergents. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2013 相似文献
16.
Endo-β-1,4-glucanase encoded byBacillus subtilis JA18 was expressed inEscherichia coli. The recombinant enzyme was purified and characterized. The purified enzyme showed a single band of 50 kDa by SDS-PAGE. The optimum pH and temperature for this endo-β-1,4-glucanase was pH 5.8 and 60 °C. The endo-β-1,4-glucanase was highly stable in a wide pH range, from 4.0 to 12.0. Furthermore, it remained stable up to 60 °C. The endo-β-1,4-glucanase was completely inhibited by 2 mM Zn2+, Cu2+, Fe3+, Ag+, whereas it is activated in the presence of Co2+. In addition, the enzyme activity was inhibited by 1 mM Mn2+ but stimulated by 10 mM Mn2+. At 1% concentration, SDS completely inhibited the enzyme. The enzyme hydrolysed carboxymethylcellulose, lichenan but no activity was detected with regard to avicel, xylan, chitosan and laminarin. For carboxymethylcellulose, the enzyme had a Km of 14.7 mg/ml. 相似文献
17.
Mohsen Mohamed Selim Asker Mohamed Fawzy Ramadan Samir Khalf Abd El-Aal Ebtsam Mokhtar Mohamed El-Kady 《World journal of microbiology & biotechnology》2009,25(5):789-794
Trehalose synthase (TSII) from Corynebacterium nitrilophilus NRC was successively purified by ammonium sulphate precipitation, ion exchange chromatography on DEAE-cellulose and gel filtration
chromatography on Sephadex G-100 columns. The specific activity of the trehalose synthase was increased ~200-fold, from 0.14
U mg−1 protein to 28.3 U mg−1 protein. TSII was found to be a monomeric protein with a molecular weight of 67–69 kDa. Characterization of the enzyme exhibited
optimum pH and temperature were 7.5 and 35°C, respectively. The purified enzyme was stable from pH 6.6 to 7.8 and able to
prolong its thermal stability up to 35°C. The enzyme activity was inhibited strongly by Zn2+, Hg2+ and Cu2+ and moderately by Ba2+, Fe2+, Pb2+ and Ni2+. Other metal ions Ca2+, Mg2+, Co2+, Mn2+ and EDTA had almost no effect. 相似文献
18.
Yu Ding Jin-Lai Miao Quan-Fu Wang Zhou Zheng Guang-You Li Ji-Chang Jian Zao-He Wu 《Polar Biology》2007,31(1):23-30
A psychrophilic glutathione reductase from Antarctic ice microalgae Chlamydomonas sp. Strain ICE-L was purified by ammonium sulfate fractionation and three steps of chromatography. The yield was up to 25.1%
of total glutathione reductase in the crude enzyme extract. The glutathione reductase activity was characterized by the spectrophotometric
method under different conditions. Purified glutathione reductase was separated by SDS-PAGE, which furnished a homogeneous
band. The native molecular mass of the enzyme was 115 kDa. Apparent Km values for NADPH and NADH (both at 0.5 mmol L−1 oxidized glutathione) were 22.3 and 83.8 μmol L−1, respectively. It was optimally active at pH 7.5, and it was stable from pH 5 to 9. Its optimum temperature was 25°C, with
activity at 0°C 23.5% of the maximum. Its optimum ion strength and optimum Mg2+ were 50–90 and 7.5 mmol L−1, respectively. Ca2+, Mg2+, and cysteine substantially increased the activity of the enzyme but chelating agents, heavy metals (Cd2+, Pb2+, Cu2+, Zn2+, etc.), NADPH, and ADP had significant inhibitory effects. This glutathione reductase can be used to study the adaptation
and mechanism of catalysis of psychrophilic enzymes, and it has a high potential as an environmental biochemical indicator
under extreme conditions. 相似文献
19.
Jeyaraman Vikramathithan Gali Nirmal kumar Pandurangan Muthuraman Kotteazeth Srikumar 《The protein journal》2010,29(7):481-486
A thermo stable xylanase was purified and characterized from the cladodes of Cereus pterogonus plant species. The enzyme was purified to homogeneity by ammonium sulfate (80%) fractionation, ion exchange and size exclusion
chromatography. The enzyme showed a final specific activity of 216.2 U/mg and the molecular mass of the protein was 80 KDa.
The optimum pH and temperature for xylanase activity were 5.0 and 80 °C, respectively,. With oat spelt xylan as a substrate
the enzyme yielded a Km value of 2.24 mg/mL and a Vmax of 5.8 μmol min−1 mg−1. In the presence of metal ions (1 mM) such as Co2+,Mn2+, Ni2+, Ca2+ and Fe3+ the activity of the enzyme increased, where as strong inhibition of the enzyme activity was observed with the use of Hg2+, Cd2+, Cu2+, while partial inhibition was noted with Zn2+ and Mg2+. The substrate specificity of the xylanase yielded maximum activity with oat spelt xylan. 相似文献
20.
《Process Biochemistry》2010,45(5):694-699
An extracellular halophilic α-amylase from Nesterenkonia sp. strain F was purified to homogeneity by 80% ethanol precipitation, Q-Sepharose anion exchange and Sephacryl S-200 gel filtration chromatography, with a 10.8-fold increase in specific activity. The molecular mass of the amylase was estimated to be 100 kDa and 106 kDa by SDS–PAGE and gel filtration chromatography, respectively. The enzyme showed maximal activity at pH 7.5 and 45 °C. The amylase was active in a wide range of salt concentrations (0–4 M) with its maximum activity at 0.5 M NaCl or 1 M KCl and was stable at the salts concentrations between 1 M and 4 M. Fe3+, Cu2+, Zn2+ and Al3+ strongly inhibited the enzyme, whereas Ca2+ stimulated the amylase activity. The α-amylase was inhibited by EDTA, but was not inhibited by PMSF and β-mercaptoethanol. The enzyme showed remarkable stability towards 0.5% SDS and sarcosyl, and 2% each of Triton X-100, Tween 80 and Tween 20. Km value of the amylase for soluble starch was 4.5 mg/ml. The amylase hydrolyzed 38% of raw wheat starch and 20% of corn starch in a period of 48 h. The major products of soluble starch hydrolysis were maltose, maltotriose and maltotetraose, indicating an α-amylase activity. 相似文献