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1.
M. Vidyasagar S. Prakash S. K. Jayalakshmi K. Sreeramulu 《World journal of microbiology & biotechnology》2007,23(5):655-662
An extremely halophilic Chromohalobacter sp. TVSP101 was isolated from solar salterns and screened for the production of extracellular halothermophilic protease.
Identification of the bacterium was done based upon biochemical tests and the 16S rRNA sequence. The partially purified enzyme
displayed maximum activity at pH 8 and required 4.5 M of NaCl for optimum proteolytic activity. In addition, this enzyme was
thermophilic and active in broad range of temperature 60–80°C with 80°C as optimum. The Chromohalobacter sp. required 4 M NaCl for its optimum growth and protease secretion and no growth was observed below 1 M of NaCl. The initial
pH of the medium for growth and enzyme production was in the range 7.0–8.0 with optimum at pH 7.2. Various cations at 1 mM
concentration in the growth medium had no significant effect in enhancing the growth and enzyme production but 0.5 M MgCl2 concentration enhanced enzyme production. Casein or skim milk powder 1% (w/v) along with 1% peptone proved to be the best
nitrogen sources for maximum biomass and enzyme production. The carbon sources glucose and glycerol repressed the protease
secretion. Immobilization of whole cells in absence of NaCl proved to be useful for continuous production of halophilic protease. 相似文献
2.
Soror SH Verma V Rao R Rasool S Koul S Qazi GN Cullum J 《Journal of industrial microbiology & biotechnology》2007,34(8):525-531
The genome sequence of Streptomyces coelicolor A3(2) contains 51 putative lipase and esterase genes mostly of unknown function. The gene estB (locus SCO 6966) was expressed as a His-tagged protein in E. coli. Esterase B was active at low temperatures exerting its maximum activity at 30°C and retaining more than 25% of its activity
at 4°C. The optimum pH was 8–8.5. The enzyme was active against short synthetic p-nitrophenylesters (C2–C10) with maximum activity towards the acetate ester (C2). The esterase was tested on 13 series of racemic esters of potential interest for the synthesis of chiral pharmaceutical
compounds. 4 of the series were substrates and a modest degree of enantioselectivity was observed (enantiomeric ratios of
1.1–1.9). 相似文献
3.
Chengtao Wang Yanping Cao Baoguo Sun Baoping Ji M. J. Robert Nout Ji Wang Yonghuan Zhao 《World journal of microbiology & biotechnology》2008,24(10):2149-2157
Rhodococcus sp. R14-2, isolated from Chinese Jin-hua ham, produces a novel extracellular cholesterol oxidase (COX). The enzyme was extracted from
fermentation broth and purified 53.1-fold based on specific activity. The purified enzyme shows a single polypeptide band
on SDS-PAGE with an estimated molecular weight of about 60 kDa, and has a pI of 8.5. The first 10 amino acid residues of the NH2-terminal sequence of the enzyme are A-P-P-V-A-S-C-R-Y-C, which differs from other known COXs. The enzyme is stable over a
rather wide pH range of 4.0–10.0. The optimum pH and temperature of the COX are pH 7.0 and 50°C, respectively. The COX rapidly
oxidizes 3β-hydroxysteroids such as cholesterol and phytosterols, but is inert toward 3α-hydroxysteroids. Thus, the presence
of a 3β-hydroxyl group appears to be essential for substrate activity. The Michaelis constant (Km) for cholesterol is estimated
at 55 μM; the COX activity was markedly inhibited by metal ions such as Hg2+ and Fe3+ and inhibitors such as p-chloromercuric benzoate, mercaptoethanol and fenpropimorph. Inhibition caused by p-chloromercuric benzoate, mercuric chloride, or silver nitrate was almost completely prevented by the addition of glutathione.
These suggests that -SH groups may be involved in the catalytic activity of the present COX. 相似文献
4.
Sudathip Titapoka Suttipun Keawsompong Dietmar Haltrich Sunee Nitisinprasert 《World journal of microbiology & biotechnology》2008,24(8):1425-1433
In order to select bacterial strains effectively secreting mannanase activity for the production of prebiotic mannooligosaccharides,
a two-step screening procedure was performed. Enriched cultures from isolation medium containing copra meal were primary screened
on an isolation agar medium containing 1% locust bean gum (LBG), which resulted in 48 mannanase-producing bacterial isolates
with significant clearing zones on the mannan-containing agar. However, only nine isolates showed appreciable mannanase activities
against copra meal in their culture supernatants (0.054–0.185 U/mg of protein) as determined in a standard assay based on
the detection of reducing sugars released from this substrate. The isolates CW2-3 and ST1-1 displayed the highest activity
against LBG and copra meal, respectively. Copra mannan hydrolysates that were obtained by using crude mannanase from these
nine isolates were further used for a secondary screening towards a growth-enhancing activity on Lactobacillus reuteri and inhibitory activity against Escherichia coli as well as Salmonella Enteritidis, resulting in 0.09–2.15 log CFU/ml enhancing activity and low inhibitory activity of 0.46–1.78 log CFU/ml as
well as 0.37–1.72 log CFU/ml, respectively. The hydrolysate of CW2-3 mannanase showed the highest enhancing activity of 2.15 log
CFU/ml while isolate ST1-1 was most effective with respect to growth inhibition against E. coli E010 and S. Enteritidis S003 with 0.76 and 1.61 log CFU/ml, respectively. Based on morphological, physical, biochemical and genetics
properties, isolates CW2-3 and ST1-1 were identified as Klebsiella oxytoca and Acinetobacter sp., respectively. Crude mannanase activity from these two strains was characterized preliminarily. The pH optima of mannanase
activity from Klebsiella oxytoca CW2-3 and Acinetobacter sp. ST1-1 were 7 and 6, respectively. The enzymes were stable at 4°C over a pH range of 3–6 and 3–10, respectively. 相似文献
5.
Gulelat D. Haki Alfredo J. Anceno Sudip K. Rakshit 《World journal of microbiology & biotechnology》2008,24(11):2517-2524
Bacillus sp. GRE1 isolated from an Ethiopian hyperthermal spring produced raw-starch digesting, Ca2+-independent thermostable α-amylase. Enzyme production in shake flask experiments using optimum nutrient supplements and environmental
conditions was 2,360 U l−1. Gel filtration chromatography yielded a purification factor of 33.6-fold and a recovery of 46.5%. The apparent molecular
weight of the enzyme was 55 kDa as determined by SDS-PAGE. Presence or absence of Ca2+ produced similar temperature optima of 65–70°C. The optimum pH was in the range of 5.5–6.0. The enzyme maintained 50% of
its original activity after 45 min of incubation at 80°C and was stable at a pH range of 5.0–9.0. The V
max and K
m values for soluble starch were 42 mg reducing sugar min−1 and 4.98 mg starch ml−1, respectively. Strong inhibitors of enzyme activity included Cu2+, Zn2+ and Fe2+. The enzyme coding gene and the deduced protein translation revealed a characteristic but markedly atypical homology to Bacillus species α-amylase sequences. The enzyme hydrolyzed wheat, corn and tapioca starch granules efficiently below their gelatinization
temperatures. Rather than the higher oligosaccharides normally produced by Bacillus α-amylases operating at high temperatures, maltose was the major hydrolysis product with the present enzyme. 相似文献
6.
The differences in pigment levels, photosynthetic activity and the chlorophyll fluorescence decrease ratio R
Fd (as indicator of photosynthetic rates) of green sun and shade leaves of three broadleaf trees (Platanus acerifolia Willd., Populus alba L., Tilia cordata Mill.) were compared. Sun leaves were characterized by higher levels of total chlorophylls a + b and total carotenoids x + c as well as higher values for the weight ratio chlorophyll (Chl) a/b (sun leaves 3.23–3.45; shade leaves: 2.74–2.81), and lower values for the ratio chlorophylls to carotenoids (a + b)/(x + c) (with 4.44–4.70 in sun leaves and 5.04–5.72 in shade leaves). Sun leaves exhibited higher photosynthetic rates P
N on a leaf area basis (mean of 9.1–10.1 μmol CO2 m−2 s−1) and Chl basis, which correlated well with the higher values of stomatal conductance G
s (range 105–180 mmol m−2 s−1), as compared to shade leaves (G
s range 25–77 mmol m−2 s−1; P
N: 3.2–3.7 μmol CO2 m−2 s−1). The higher photosynthetic rates could also be detected via imaging the Chl fluorescence decrease ratio R
Fd, which possessed higher values in sun leaves (2.8–3.0) as compared to shade leaves (1.4–1.8). In addition, via R
Fd images it was shown that the photosynthetic activity of the leaves of all trees exhibits a large heterogeneity across the
leaf area, and in general to a higher extent in sun leaves than in shade leaves. 相似文献
7.
C. K. Deepa Syed G. Dastager Ashok Pandey 《World journal of microbiology & biotechnology》2010,26(7):1233-1240
The efficacy of four potential phosphate solubilizing Enterobacter isolated from non-rhizospheric soil in Western ghat forest in India. Plant growth promoting ability of these isolates was evaluated in cowpea. All are gram negative, rod shaped,
0.8–1.6 mm in size, and psychrotrophic in nature, grow from 5 to 40°C (optimum temp. 28 ± 2°C). All isolates exhibits growth
at a wide range of pH 6–12, optimum at pH 7.0 and tolerates up to 7% (w/v) salt concentration. 16S rRNA gene sequencing reveals
the confirmation of isolates to Enterobacter aerogenes sp. (NII-0907 and NII-0929), Enterobacter cloacae subsp. cloacae sp. (NII-0931) and Enterobacter asburiae sp. (NII-0934) with which they share >99% sequence similarity. Under in vitro conditions, all the four isolates were found
to produce indole acetic acid, P-solubilization and hydrogen cyanide. The P-solubilizing activity coincided with a concomitant
decrease in pH of the medium (pH 7.0–<3.0). The plant growth promotion properties were demonstrated through a cow pea (Vigna unguiculata (L.) walp) based bioassay under greenhouse conditions. Although the bacterial inoculation was found to result in significant increment
in root, shoot and biomass and it stimulated bacterial counts in the rhizosphere. Hence, these isolates can further formulated
and used for field application. 相似文献
8.
Damayanti De Ananda Mukhopadhyay Ranadhir Chakraborty 《World journal of microbiology & biotechnology》2008,24(11):2727-2729
Enterobacter sp. was isolated from the diseased and dead caterpillars of the tea leaf roller (Caloptilia theivora) from the Darjeeling foothill region. When the vegetative form of the bacterium was applied via food, mortality of C. theivora showed an LC50 value at 363.1 μg/ml (bacterial wt./vol. of water) with fiducial limits 363.25 and 362.94 μg/ml respectively. The LT50 values for C. theivora were 6 days for 100 μg/ml, 5.96 days for 300 μg/ml, 5.81 days for 500 μg/ml, 4.96 days for 750 μg/ml and 4.61 days for 1,000 μg/ml
concentrations. The finding would enable one to contemplate development of a microbial pesticide using this novel Enterobacter sp. DD01 for control of the leaf rolling pest. 相似文献
9.
The importance of various parameters such as sugarcane juice concentration, pH of the medium, and effects of different solid
supports for maximum secretion of pectin lyase from Penicillium citrinum MTCC 8897 has been studied. The enzyme was purified to homogeneity by Sephadex G-100 and DEAE-cellulose chromatography. The
molecular mass determined by SDS-PAGE was 31 kDa. The K
m and k
cat values were found to be 1 mg/ml and 76 sec−1, respectively. The optimum pH of the purified pectin lyase was 9.0, though it retains activity in the pH 9.0–12.0 range when
exposed for 24 h. The optimum temperature was 50°C, and the pectin lyase was found to be completely stable up to 40°C when
exposed for 1 h. The purified pectin lyase was found efficient in retting of Linum usitatissimum, Cannabis sativa, and Crotalaria juncea.
Published in Russian in Biokhimiya, 2009, Vol. 74, No. 7, pp. 985–992. 相似文献
10.
M. Salam N. Dahiya R. Sharma S. K. Soni G. S. Hoondal R. Tewari 《Indian journal of microbiology》2008,48(3):358-364
A chitinase producing bacterium Enterobacter sp. NRG4, previously isolated in our laboratory, has been reported to have a wide range of applications such as anti-fungal
activity, generation of fungal protoplasts and production of chitobiose and N-acetyl D-glucosamine from swollen chitin. In
this paper, the gene coding for Enterobacter chitinase has been cloned and expressed in Escherichia coli BL21(DE3). The structural portion of the chitinase gene comprised of 1686 bp. The deduced amino acid sequence of chitinase
has high degree of homology (99.0%) with chitinase from Serratia marcescens. The recombinant chitinase was purified to near homogeneity using His-Tag affinity chromatography. The purified recombinant
chitinase had a specific activity of 2041.6 U mg−1. It exhibited similar properties pH and temperature optima of 5.5 and 45°C respectively as that of native chitinase. Using
swollen chitin as a substrate, the Km, kcat and catalytic efficiency (kcat/Km) values of recombinant chitinase were found to be 1.27 mg ml−1, 0.69 s−1 and 0.54 s−1M−1 respectively. Like native chitinase, the recombinant chitinase produced medicinally important N-acetyl D-glucosamine and
chitobiose from swollen chitin and also inhibited the growth of many fungi. 相似文献
11.
The chemolithoautotroph, Arthrobacter sp.15b oxidizes arsenite to arsenate using a membrane bound arsenite oxidase. The enzyme arsenite oxidase is purified to
its homogeneity and identified using MALDI-TOF MS analysis. Upon further characterization, it was observed that the enzyme
is a heterodimer showing native molecular mass as ~100 kDa and appeared as two subunits of ~85 kDa LSU and 14 kDa SSU on SDS–PAGE.
The V
max and K
m values of the enzyme was found to be 2.45 μM (AsIII)/min/mg) and 26 μM, respectively. The purified enzyme could withstand
wide range of pH and temperature changes. The enzyme, however, gets deactivated in the presence of 1 mM of DEPC suggesting
the involvement of histidine at the binding site of the enzyme. The peptide analysis of large sub unit of the enzyme showed
close match with the arsenite oxidases of Burkholderia sp. YI019A and arsenite oxidase, Mo-pterin containing subunit of Alcaligenes
faecalis. The small subunit, however, differed from other arsenite oxidases and matched only with 2Fe–2S binding protein of Anaplasma
phagocytophilum. This indicates that Rieske subunits containing the iron–sulfur clusters present in the large as well as small subunits of
the enzyme are integral part of the protein. 相似文献
12.
Jeyagowri Kiddinamoorthy Alfredo J. Anceno Gulelat D. Haki Sudip K. Rakshit 《World journal of microbiology & biotechnology》2008,24(5):605-612
Production of extracellular xylanase from Bacillus sp. GRE7 using a bench-top bioreactor and solid-state fermentation (SSF) was attempted. SSF using wheat bran as substrate
and submerged cultivation using oat-spelt xylan as substrate resulted in an enzyme productivity of 3,950 IU g−1 bran and 180 IU ml−1, respectively. The purified enzyme had an apparent molecular weight of 42 kDa and showed optimum activity at 70°C and pH
7. The enzyme was stable at 60–80°C at pH 7 and pH 5–11 at 37°C. Metal ions Mn2+ and Co2+ increased activity by twofold, while Cu2+ and Fe2+ reduced activity by fivefold as compared to the control. At 60°C and pH 6, the K
m for oat-spelt xylan was 2.23 mg ml−1 and V
max was 296.8 IU mg−1 protein. In the enzymatic prebleaching of eucalyptus Kraft pulp, the release of chromophores, formation of reducing sugars
and brightness was higher while the Kappa number was lower than the control with increased enzyme dosage at 30% reduction
of the original chlorine dioxide usage. The thermostability, alkali-tolerance, negligible presence of cellulolytic activity,
ability to improve brightness and capacity to reduce chlorine dioxide usage demonstrates the high potential of the enzyme
for application in the biobleaching of Kraft pulp. 相似文献
13.
Prangya Paramita Tripathy S. Ayyappan 《World journal of microbiology & biotechnology》2005,21(8-9):1339-1343
Summary A preliminary comparative evaluation of the two commonly encountered free-living nitrogen fixers in the aquatic system, Azotobacter and Azospirillum was carried out in the laboratory for use as biofertilizers in aquaculture considering the importance of eco-friendly and
econo-friendly productivity optimization of freshwater aquaculture. The ammonium–nitrogen levels in water media in Azotobacter treatment varied in the range 2.59–34.34 μg-N/l and was found to be significantly different from that of Azospirillum treatment (p < 0.05). The viable population of the respective nitrogen fixers as colony forming units (c.f.u.) in water media in charcoal-immobilized
Azotobacter treatment ranged from 0.39 to 2.48 × 103 c.f.u./ml and were significantly higher (p < 0.05) than the counterparts. The nitrogenase activity in the same treatment similarly remained higher, at 8.3–12.15 nmol
of ethylene/ml water/h followed by the alginate-immobilized Azotobacter treatment which was at 7.2–11.40 nmol of ethylene/ml water/h compared to 5.8–7.8 and 4.65–4.83 in the respective Azospirillum-treated counterparts. Hence, better performance of Azotobacter sp. over Azospirillum sp., and of charcoal immobilization over alginate immobilization were observed. 相似文献
14.
6-Phosphogluconate dehydrogenase (6PG) was purified from rat small intestine with 36% yield and a specific activity of 15 U/mg.
On SDS/PAGE, one band with a mass of 52 kDa was found. On native PAGE three protein and two activity bands were observed.
The pH optimum was 7.35. Using Arrhenius plots, Ea, ΔH, Q10 and Tm for 6PGD were found to be 7.52 kcal/mol, 6.90 kcal/mol, 1.49 and 49.4°C, respectively. The enzyme obeyed “Rapid Equilibrium
Random Bi Bi” kinetic model with Km values of 595 ± 213 μM for 6PG and 53.03±1.99 μM for NADP. 1/Vm versus 1/6PG and 1/NADP plots gave a Vm value of 8.91±1.92 U/mg protein. NADPH is the competitive inhibitor with a Ki of 31.91±1.31 μM. The relatively small Ki for the 6PGD:NADPH complex indicates the importance of NADPH in the regulation of the pentose phosphate pathway through G6PD
and 6PGD. 相似文献
15.
Sorokin ID Zadorina EV Kravchenko IK Boulygina ES Tourova TP Sorokin DY 《Extremophiles : life under extreme conditions》2008,12(6):819-827
Gram-positive bacteria capable of nitrogen fixation were obtained in microoxic enrichments from soda soils in south-western
Siberia, north-eastern Mongolia, and the Lybian desert (Egypt). The same organisms were obtained in anoxic enrichments with
glucose from soda lake sediments in the Kulunda Steppe (Altai, Russia) using nitrogen-free alkaline medium of pH 10. The isolates
were represented by thin motile rods forming terminal round endospores. They are strictly fermentative saccharolytic anaerobes
but tolerate high oxygen concentrations, probably due to a high catalase activity. All of the strains are obligately alkaliphilic
and highly salt-tolerant natronophiles (chloride-independent sodaphiles). Growth was possible within a pH range from 7.5 to
10.6, with an optimum at 9.5–10, and within a salt range from 0.2 to 4 M Na+, with an optimum at 0.5–1.5 M for the different strains. The nitrogenase activity in the whole cells also had an alkaline
pH optimum but was much more sensitive to high salt concentrations compared to the growing cells. The isolates formed a compact
genetic group with a high level of DNA similarity. Phylogenetic analysis based on 16S-rRNA gene sequences placed the isolates
into Bacillus rRNA group 1 as a separate lineage with Amphibacillus tropicus as the nearest relative. In all isolates the key functional nitrogenase gene nifH was detected. A new genus and species, Natronobacillus azotifigens gen. nov., sp. nov., is proposed to accommodate the novel diazotrophic haloalkaliphiles.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
The GenBank accession numbers for the 16S rRNA gene of the novel strains are EU143681-EU143690 and EU850814-EU850816; for
the nifH gene the accession numbers are EU542601, EU563380-EU563386 and EU850817-EU850819. 相似文献
16.
Fawzeia H. Elmhalli Katinka Pålsson Jan Örberg Thomas G. T. Jaenson 《Experimental & applied acarology》2009,48(3):251-262
The toxicity of para-menthane-3,8-diol (PMD), the main arthropod-repellent compound in the oil of the lemon eucalyptus, Corymbia citriodora, was evaluated against nymphs of Ixodes ricinus using five methods (A–E) of a contact toxicity bioassay. Mortality rates were estimated by recording numbers of dead nymphs
at 30 min intervals during the first 5 h after the start of exposure and at longer intervals thereafter. The mortality rate
increased with increasing concentration of PMD and duration of exposure with a distinct effect after 3.5 h. From the results
obtained by methods A, C and E, the LC50 range was 0.035–0.037 mg PMD/cm2 and the LC95 range was 0.095–0.097 mg PMD/cm2 at 4 h of exposure; the LT50 range was 2.1–2.8 h and the LT95 range was 3.9–4.2 h at 0.1 mg PMD/cm2. To determine the duration of toxic activity of PMD, different concentrations (0.002, 0.01, 0.1 mg PMD/cm2) were tested and mortality was recorded at each concentration after 1 h; thereafter new ticks were tested. This test revealed
that the lethal activity of PMD remained for 24 h but appeared absent after 48 h. The overall results show that PMD is toxic
to nymphs of I. ricinus and may be useful for tick control. 相似文献
17.
Renata Matraszek 《Acta Physiologiae Plantarum》2008,30(3):361-370
The author studied the effect of different nickel concentrations (0, 0.4, 40 and 80 μM Ni) on the nitrate reductase (NR) activity
of New Zealand spinach (Tetragonia expansa Murr.) and lettuce (Lactuca sativa L. cv. Justyna) plants supplied with different nitrogen forms (NO3
−–N, NH4
+–N, NH4NO3). A low concentration of Ni (0.4 μM) did not cause statistically significant changes of the nitrate reductase activity in
lettuce plants supplied with nitrate nitrogen (NO3
−–N) or mixed (NH4NO3) nitrogen form, but in New Zealand spinach leaves the enzyme activity decreased and increased, respectively. The introduction
of 0.4 μM Ni in the medium containing ammonium ions as a sole source of nitrogen resulted in significantly increased NR activity
in lettuce roots, and did not cause statistically significant changes of the enzyme activity in New Zealand spinach plants.
At a high nickel level (Ni 40 or 80 μM), a significant decrease in the NR activity was observed in New Zealand spinach plants
treated with nitrate or mixed nitrogen form, but it was much more marked in leaves than in roots. An exception was lack of
significant changes of the enzyme activity in spinach leaves when plants were treated with 40 μM Ni and supplied with mixed
nitrogen form, which resulted in the stronger reduction of the enzyme activity in roots than in leaves. The statistically
significant drop in the NR activity was recorded in the aboveground parts of nickel-stressed lettuce plants supplied with
NO3
−–N or NH4NO3. At the same time, there were no statistically significant changes recorded in lettuce roots, except for the drop of the
enzyme activity in the roots of NO3
−-fed plants grown in the nutrient solution containing 80 μM Ni. An addition of high nickel doses to the nutrient solution
contained ammonium nitrogen (NH4
+–N) did not affect the NR activity in New Zealand spinach plants and caused a high increase of this enzyme in lettuce organs,
especially in roots. It should be stressed that, independently of nickel dose in New Zealand spinach plants supplied with
ammonium form, NR activity in roots was dramatically higher than that in leaves. Moreover, in New Zealand spinach plants treated
with NH4
+–N the enzyme activity in roots was even higher than in those supplied with NO3
−–N. 相似文献
18.
An endopolygalacturonase of Rhizopus sp. strain LKN, one of several isolates from tempe starter (ragi), was purified 235-fold by CM-Sephadex C-50, DEAE-Sephadex A-50 ion exchange chromatographies and Sephadex G-75 gel filtration. The purified enzyme was homogeneous by SDS-PAGE with a M
r of 38.5 kDa. Its K
m value for pectic acid was 2 mg/ml. It was stable at pH 4.5 to 11 and up to 50°C, with optimum activity at pH 4.5 to 4.75 and 55 to 60°C. Some ionic compounds enhanced the enzyme activity, whereas tannic acid at 0.5 mm caused about 90% inhibition.The authors are with the Department of Food Science and Technology, Faculty of Agriculture, Kyushu University, Hakozaki, Fukuoka 812, Japan. 相似文献
19.
Nack-Shick Choi Jae Jun Song Dong-Min Chung Young Jae Kim Pil Jae Maeng Seung-Ho Kim 《Journal of industrial microbiology & biotechnology》2009,36(3):417-426
A novel fibrinolytic enzyme (AJ) was purified from Staphylococcus sp. strain AJ screened from Korean salt-fermented Anchovy-jeot. Relative molecular weight of AJ was determined as 26 kDa by using SDS-PAGE and fibrin zymography. Based on a 2D gel, AJ
was found to consist of three active isoforms (pI 5.5–6.0) with the same N-terminal amino acid sequence. AJ exhibited optimum pH and temperature at 2.5–3.0 and 85°C, respectively.
AJ kept 85% of the initial activity after heating at 100°C for 20 min on the zymogram gel. The Michaelis constant (K
m) and K
cat values of AJ towards α-casein were 0.38 mM and 19.73 s−1, respectively. AJ cleaved the Aα-chain of fibrinogen but did not affect the Bβ- and γ-chains, indicating that it is an α-fibrinogenase.
The fibrinolytic activity was inhibited by diisopropyl fluorophosphate, indicating AJ is a serine protease. Interestingly,
AJ was very stable at acidic condition, SDS, and heat (100°C), whereas it was easily degraded at neutral and alkaline conditions.
In particular, AJ formed an active homo-dimer in the pH range from 7.0 to 8.0. To our knowledge, a similar combination of
acid and heat stability has not yet been reported for other fibrinolytic enzymes. 相似文献
20.
The effects of phosphorus, Zn2+, CO2, and light intensity on growth, biochemical composition, and the activity of extracellular carbonic anhydrase (CA) in Isochrysis galbana were investigated. A significant change was observed when the concentration of phosphorus in the medium was increased from
5 μmol/L to 1000 μmol/L affecting I. galbana’s cell density, biochemical composition, and the activity of extracellular CA. Phosphorous concentration of 50 μmol/L to 500
μmol/L was optimal for this microalgae. The Zn2+ concentration at 10 μmol/L was essential to maintain optimal growth of the cells, but a higher concentration of Zn2+ (≥ 1000 μmol/L) inhibited the growth of I. galbana. High CO2 concentrations (43.75 mL/L) significantly increased the cell densities compared to low CO2 concentrations (0.35 mL/L). However, the activity of extracellular CA decreased significantly with an increasing concentration
of CO2. The activity of extracellular CA at a CO2 concentration of 43.75 mL/L was approximately 1/6 of the activity when the CO2 concentration was at 0.35 mL/L CO2. Light intensity from 4.0 mW/cm2 to 5.6 mW/cm2 was beneficial for the growth, biochemical composition and the activity of extracellular CA. The lower and higher light intensity
was restrictive for growth and changed its biochemical composition and the activity of extracellular CA. These results indicate
that phosphorus, Zn2+, CO2, and light intensity are important factors that impact growth, biochemical composition and the activity of extracellular
CA in I. galbana. 相似文献