Purification and characterization of cellulases from Clostridium papyrosolvens

The extracellular cellulolytic enzyme complex from Clostridium papyrosolvens was isolated from a culture of this organism grown on filter paper. The complex showed xylanase, carboxymethyl cellulase (CMCase) and Avicelase (but not β-glucosidase) activities. Non-denaturing polyacrylamide gel electrophoresis (PAGE) revealed two dominant bands, sited at the origin (corresponding to a fraction of high molecular weight) and at 280 kDa, and eight other very weak bands in the 20–180 kDa MW range. Gel overlay techniques showed strong CMCase activity in the region comprised between the origin and 320 kDa and in three distinct sites in the 40–230 kDa region. Xylanase activities were detected as a continuous band almost covering all the track. SDS—PAGE gave a multiplicity of different intensity bands in the 20–130 kDa range. Cellulolytic activities (CMCase in the 85–90 kDa range) and hemicellulolytic activities (xylanase at approximately 95, 60, 42 and 32 kDa) were detected through application of the corresponding CMC and xylan overlay techniques. Though anion-exchange chromatography (DEAE-Sephadex A-50) and gel filtration (Biogel P-100) techniques did not permit a good separation of the different cellulolytic activities, an 11.3-fold increase of the Avicelase specific activity was achieved in a fraction containing 38.5% of the original total activity. Maximum enzymatic activities in crude preparations were observed at pH 5.4 and 50°C for xylanase and at pH 4.8 and 45°C for CMCase. The Michaelis constants for CMCase and xylanase were respectively: V_max = 9.1 μg glucose equivalents min^? ml^?, Km=3.3 g CMcellulose liter^? and V_max=44.5 μg xylose equiuulents min^? ml^? and K_m=2.7 g xylun liter^?. Both enzymes were inhibited by a competitive mechanism.     

Saccharification of microcrystalline cellulose by cellulase of Morchella conica mushroom mycelium in comparison with a commercial cellulase

A comparison was made of the hydrolysis of microcrystalline cellulose Avicel using the cellulolytic complex of the ascomycete Morchella conica and a commercial cellulase. An enzyme concentration of 1 U AVase mg^?1 substrate gave the best results during the hydrolytic processes. At 24 h, the M. conica enzyme complex achieved 35.5% and the commercial cellulase 31·2% saccharification, with glucose 84·2% and 52% of the total reducing sugars liberated, respectively. The specific rates of hydrolyses were 0·77 and 0·14 h^?1 for reducing sugars and 0·54 and 0·12 h^?1 for glucose formation with M. conica and the commercial cellulase, respectively. At 96 h, the degree of saccharification reached 46% for M. conica and 49% for the commercial cellulase, with glucose 76·5% and 65·9% of the total reducing sugars liberated, respectively. Both the complexes were quite stable with a residual activity of 62% CMCase and 47% AVase for M. conica, and 74% CMCase and 57% AVase for the commercial cellulase at 96 h of hydrolysis. The qualitative analysis of the hydrolysis products by TLC indicated, for M. conica, an earlier appearance of cellobiose, which was quickly hydrolyzed to glucose.     

Optimization of salts in medium for production of carboxymethylcellulase by a psychrophilic marine bacterium, <Emphasis Type="Italic">Psychrobacter aquimaris</Emphasis> LBH-10 using two statistical methods

The concentration of four mineral salts in the medium for the production of carboxymethylcellulase (CMCase) by Psychrobacter aquimaris LBH-10 were optimized using orthogonal array method (OAM) and response surface method (RSM) and their results from two statistical methods were compared. The analysis of variance (ANOVA) of data from central composite design (CCD) based on OAM indicated that potassium phosphate gave the highest percentage participation for cell growth as well as production of CMCase. However, their relative participations of four salts for cell growth were different from those for production of CMCase. The ANOVA of results from RSM indicated that highly significant factors (“probe>F” less than 0.0001) for cell growth were K₂HPO₄ and (NH₄)₂SO₄, whereas those for production of CMCase were K₂HPO₄, NaCl, and MgSO₄·7H₂O. The optimal concentration of K₂HPO₄, NaCl, MgSO₄·7H₂O, and (NH₄)₂SO₄ for cell growth extracted by Design Expert Software based on RSM were 7.10, 0.84, 0.24, and 0.33 g/L, respectively, whereas those for production of CMCase were 3.00, 0.52, 0.34, and 0.45 g/L. The optimal concentrations of salts for cell growth and production of CMCase using RSM basically coincided with those using OAM as well as those from ‘one-factor-at-a-time’ method. The production of CMCase by P. aquimaris LBH-10 with optimized concentrations of salts was 273.0 U/mL, which was enhanced by 1.27 times higher than the previous report.     

Cellulolytic properties of Chaetomium crispatum

The cellulolytic properties of a Chaetomium crispatum strain were investigated. The cellulolytic enzyme complex i.e.: exo-1,4-β-glucosidase (EC 3.2.1.74); endo-1,4-β-glucanase (EC 3.2.1.4.) and β-glucosidase or cellobiase (EC 3.2.1.21) displayed optimal activity at pH 5.0 and 25°C. Although carboxymethyl-celluloses are the usual pseudo-substrates for this enzyme complex, those with a high degree of substitution gave rise to poor growth and low cellulase activity. Insoluble crude cellulosics such as newsprint, recycled paper, rice and flax straw were substantially solubilised at 28°C within 3–5 days of fermentation. A study of the cellulase-complex formation during the growth cycle revealed that β-glucosidase was produced mainly intracellularly in the early exponential phase, while the overall exo-1,4-β-glucosidase and endo-1,4-β-glucanase formation gradually increased during the total fermentation cycle. The mycelial protein of Chaetomium crispatum grown on crude cellulosics displayed a favourable amino acid pattern, indicating its potential value as a source of single cell protein (SCP).     

Vitamin E. Deficiency increases the synthesis of platelet-activating factor (PAF) in rat polymorphonuclear leucocytes

Vitamin E deficiency was found to stimulate FMLP (N-formyl-L-methionyl-L-leucyl-L-phenylalanine)-induced biosynthesis of PAF (1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) in polymorphonuclear leucocytes (PMN) from rat peritoneum. In three separate experiments each, the amounts of PAF synthesized during 6min and 12 min incubation of PMN cells from control, vitamin E-supplemented, and vitamin E-deficient rats were 129–240, 131–227 and 248–354 pmol/10⁶ cells, respectively. The activity of the acetyl-transferase, which transfers the acetyl moiety of [³H]acetyl-CoA to 2-lysoPAF (1-O-alkyl-sn-glycero-3-phosphocholine) to form [³H]PAF, was higher in PMN homogenates from vitamin E-deficient rats (2.28±0.07 nmol/min/mg protein) than in those from E-supplemented rats (1.06±0.10 nmol/min/mg protein). However, there was no difference between the two groups in the activity of acetylhydrolase (4.26±0.71 and 4.26±0.06 nmol/min/mg protein, respectively), measured as degradation of [³H]PAF to [³H]lysoPAF.In vitro addition of α-tocopherol did not inhibit the increased activity of acetyl-transferase in vitamin E-deficient rats, in-dicating that the enzyme in vitamin E-supplemented rats was not directly inhibited by α-tocopherol. The acetyltransferases of the two groups showed similar Km values for acetyl-CoA, but different Vmax values (225 μM and 6.4 nmol/min/mg protein in vitamin E-deficient rats, and 216 μM and 3.6 nmol/min/mg protein in vitamin E-supplemented rats), suggesting that the enzyme was not activated but increased in amount in vitamin E deficiency.     

Purification and characterization of an extracellular lipase from Penicillium candidum

Penicillium candidum produces and secretes a single extracellular lipase with a monomer molecular weight of 29 kDa. However, this enzyme forms dimers and higher molecular weight aggregates under nondenaturing conditions. The lipase from P. candidum was purified 37-fold using Octyl-Sepharose CL-4B and DEAE-Sephadex columns. The optimal assay conditions for lipase activity were 35°C and pH 9. The lipase was stable in the pH range of 5–6 with a pl of 5.5, but rapid loss of the enzyme activity was observed above 25°C. Tributyrin was found to be the best substrate for the P. candidum lipase, among those tested. Metal ions such as Fe²⁺ and Cu²⁺ inhibited enzymatic activity and only Ca²⁺ was able to slightly enhance lipase activity. Ionic detergents inhibited the activity of the enzyme, whereas nonionic detergents stimulated lipase activity.     

Xylanase production by Bacillus polymyxa

Bacillus polymyxa produced high levels (12–13 U cm^?3) of extracellular xylanases when grown in a complex medium containing yeast extract and oat spelt xylan as nitrogen and carbon sources respectively. Substantially lower yields of enzyme were produced during growth on the monosaccharides glucose, arabinose and xylose. Meagre growth occurred when ammonium sulphate, instead of yeast extract, was used as nitrogen source. When assayed in culture broth supernatants, xylanase showed an optimum activity in 48°C and at pH values in the range 5.0–6.5. Under such conditions, the half-life of this xylanase preparation was 8 h. Mn²⁺ showed a strong inhibitory effect on the enzyme, but inhibition by EDTA (27% w/v) suggested dependence on a metallic ion. SDS-PAGE and zymogram overlay showed that up to five separate xylanases in the range of 20 to 116 kDa were produced.     

Purification and characterization of a 28 kDa cytosolic inhibitor of cholesteryl ester hydrolases in rat testis

A 28 kDa inhibitory protein was purified from ratestis cytosol by sequential 40–65% ammonium sulfate precipitation, cation exchange chromatography, anion exchange chromatography, and preparative SDS-polyacrylamide gel electrophoresis. The heat-stable, trypsin-labile protein exhibited nonenzymatic, concentration-dependent inhibition of testicular and pancreatic cholesteryl ester hydrolases at all stages of putification. Copurifying at each stage was a 26.5 kDa protein which comprised 25% of the mass of the two proteins. Polyclonal antibodies raised to either or both 28 kDa and 26.5 kDa proteins by direct injection of excised electrophoretic bands cross-reacted with both proteins on western blots, immunoprecipitated both proteins, and neutralized inhibitory activity. Amino acid compositions of the individual proteins electroeluted from SDS-polyacrylamide gels were different from those of other surface-active proteins of similar molecular weights. Both proteins exhibited identical pl of 4.8 on chromatofocusing columns and two-dimensional gel electrophoresis. Although the subcellular distribution of the 28 kDa protein is unknown, its testicular cytosolic concentration, calculated from the purified protein mass, was 8×10⁻⁹ mols/L, which probably underestimates the actual concentration by an order of magnitude. This is greater than the minimum concentration required forin vitro inhibition (10⁻⁹ mols/L), consistent with a physiological role for this protein.     

Purification and enzymatic properties of the extracellular and constitutive chitosanase produced by Bacillus subtilis RKY3

BACKGROUND: Purification and enzymatic properties of a chitosanase from Bacillus subtilis RKY3 have been investigated to produce a chitooligosaccharide. The enzyme reported was extracellular and constitutive, which was purified by two sequential steps including ammonium sulfate precipitation and ion exchange chromatography. RESULTS: Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis of the purified chitosanase revealed one single band corresponding to a molecular weight of around 24 kDa. The highest chitosanase activity was found to be at pH 6.0 and at 60 °C. Although the mercaptide forming agents such as Hg²⁺ (10 mmol L^?1) and p‐hydroxymercuribenzoic acid (1 mmol L^?1, 10 mmol L^?1) significantly or totally inhibited the enzyme activity, its activity was enhanced by the presence of 10 mmol L^?1 Mn²⁺. The enzyme showed activity for hydrolysis of soluble chitosan and glycol chitosan, but colloidal chitin, carboxymethyl cellulose, crystalline cellulose, and soluble starch were not hydrolyzed. The analysis of chitosan hydrolysis by thin‐layer chromatography and viscosity variation revealed that the purified enzyme should be endosplitting‐type chitosanase. CONCLUSION: The chitosanase produced by Bacillus subtilis RKY3 was a novel chitosanlytic enzyme with relatively low molecular weight, which is a versatile enzyme for chitosan hydrolysis because it could hydrolyze soluble chitosan into a biofunctional oligosaccharide at a high level. Copyright © 2011 Society of Chemical Industry     

Vinyl‐type homo‐ and copolymerization of norbornene catalyzed by bis(phenoxyimine) titanium complex

A ternary catalytic system consisting of a bis(phenoxyimine) titanium complex, triisobutylaluminium and an organoboron compound exhibited high activity in the vinyl‐type homopolymerization of norbornene. The obtained polynorbornene showed a modest molecular weight (M _n ≈ 5 × 10⁴ g mol^?1) and broad molecular weight distribution (polydispersity index ≈ 3.5). A copolymer of norbornene with 1,3‐butadiene was prepared using a binary catalytic system consisting of bis(phenoxyimine) titanium complex and triisobutylaluminium. The norbornene units in the copolymer adopted a vinyl‐type addition structure confirmed using distortionless enhancement by polarization transfer 135 ¹³C NMR microstructure analyses. Polymerization kinetics studies showed that neither monomer feed ratio nor conversion had an effect on the composition of the copolymer backbone which was composed of 55% norbornene units and 45% 1,3‐butadiene units. The essentially constant polymer composition implied an alternating nature of chain propagation. The copolymer exhibited good thermal stability and moderate glass transition temperature (50.9–68.2 °C) with a relatively high molecular weight (M _w = 0.18 × 10–1.31 × 10⁵ g mol^?1), and excellent transparency (maximal transmittance >80%). © 2017 Society of Chemical Industry     

Overexpression of aRhizopus delemar lipase gene inEscherichia coli

A cloned complementary deoxyribonucleic acid encoding the precursor polypeptide of an extracellular lipase from the fungusRhizopus delemar was altered by site-directed mutagenesis to generate deoxyribonucleic acid fragments that specifically code for the polypeptides of the proenzyme and the mature form of the lipase. Attempts to produce these polypeptides in enzymatically active form inEscherichia coli revealed toxic effects toward the host. Therefore the polypeptides were expressed as inactive and insoluble forms in the cytoplasm ofE. coli BL21 (DE3) cells using plasmid vector pET11-d. With this tightly regulated high-level expression system, lipase and prolipase polypeptides were produced to estimated levels of up to 21% and 15%, respectively, of total cellular protein. The insoluble polypeptides were solubilized in 8 M urea. Refolding into active forms was achieved by treatment with the redox system cystine/cysteine and dilution. Refolded mature lipase was purified to homogeneity by affinity and ion exchange chromatography. The enzyme had a specific activity comparable to that of lipase from the fungal culture. The quantities of pure enzyme obtained from a 1-L culture ofE. coli exceeded those obtained from the fungal culture by a factor of at least 100. Refolded recombinant prolipase was purified essentially to homogeneity and had a specific activity similar to that of the mature enzyme. Its pH optimum was 7.5, rather than the pH 8 determined for recombinant mature lipase and for the enzyme purified from the fungal culture. Recombinant prolipase retained activity after 15 min incubation at 65°C, while mature lipase retained activity only up to 45°C. Mention of brand or firm names does not constitute an endorsement by the U.S. Department of Agriculture over others of a similar nature not mentioned.     

Comparison of cellulolytic enzyme complexes of different fungal origin

The cellulolytic culture filtrates of Trichoderma reesei QM 9414, Aspergillus terreus OKI 16/5 and Penicillium verruculosum WA 30 were purified and separated into components by one-step preparative isoelectric focusing (IF). The culture filtrates, the mycelia and the separated components were investigated for cellulolytic (filter-paper degrading (FPA), carboxymethylcellulose degrading (C_x), cotton hydrolyzing (C₁), cellobiase) and proteinase B activities. The molecular weights of the cellulolytic fractions were determined by SDS-electrophoresis. The culture filtrates differed in the proportions of the various cellulolytic activities. The analytical IF separation of the enzyme complexes resulted in a total of 28–31 protein fractions, mostly glycoproteins. The complexes of the different culture filtrates were separated, by preparative IF, into 28 fractions each. The greatest part of the activities could be recovered in fractions 8–18. The activities in the fractions were recovered to different extents. The main protein components in the enzyme complexes were found to be endoglucanases (C_x) with pI values between 3 and 6. Most of the endoglucanases proved to be glycoproteins. FPA and cellobiase activities were recovered to various extents in the fractions of the three culture filtrates. Only small parts of the C₁ activities were recovered in the Trichoderma and Penicillium fractions. The originally low C₁ activity of the Aspergillus culture filtrate was recovered to 76% in the separated fractions. Most of the C_x activities were found to be lost. However, the separation losses were found to be reversible: on combining the fractions, the activities originally present in the culture filtrate were nearly entirely restored. The molecular weights of the fractions of the different culture filtrates covered the range 10 000 to 70 000 D. The IF fractions of similar activity patterns also showed similar molecular weight distributions. The relatively large number of isoenzymes is assumed to be the result of the endocellular proteinase activity present, which splits off the cellulase enzyme components from proenzymes of high molecular weight during the fermentation process.     

Production of cellulases from coconut coir pith in solid state fermentation

Coconut coir pith, available in abundance especially in tropical countries, was studied as a substrate for the production of cellulase[1,4(1,3;1,4)-β-D -glucan 4-glucanohydrolase, EC 3.2.1.4] and β-D -glucosidase(β-D -glucoside glucohydrolase, EC 3.2.1.21) in solid state fermentation. The effects of fermentation time, nutrient level, substrate particle size and inoculum size have been examined for optimal production of these enzymes by the fungal strain Aspergillus niger NCIM 1005. The highest filter paper activity (FPA) of 4.11 IU g^?1, carboxyl methyl cellulose (CMCase) activity of 15·55 IU g^?1 and cellobiase activity of 9·31 IU g^?1 were obtained after 7 to 8 days of fermentation. Reese and Mandel's mineral solution in the substrate to mineral solution ratio of 1:10 (w/v) supported high cellulase and cellobiase activities. An inoculum size of 20–50% (v/v) based on the volume of mineral medium and substrate average particle size of 375 μm were optimum for enzyme production.     

Effect of lipids and surfactants on extracellular lipase production by Yarrowia lipolytica

The production of extracellular lipase in submerged cultures of Yarrowia lipolytica CECT 1240 has been investigated. Several compounds have been added to the culture medium, in order to assess their efficiency as inducers of lipase production. First, the effect of triglycerides (olive oil, sunflower oil, tributyrin) and fatty acids (oleic acid) has been studied. The highest activity level was obtained with sunflower oil (58 U cm^?3), followed by olive oil (49 U cm^?3). The cultures with tributyrin and oleic acid attained similar activities (33 U cm^?3). Then, several surfactants (Tween 80, Triton X‐100, gum arabic, polyethylene glycol 200) were added to the cultures with sunflower oil, in an attempt to increase the levels of extracellular lipase activity. The obtained activities were slightly lower than those achieved without surfactants. The assay of a wide range of surfactant concentrations in the case of PEG‐200 (with which the highest activity levels had been attained) did not improve the results. This strain secreted lipase concentrations two‐fold higher and showed significantly different behaviour towards the presence of surfactants in the culture medium, compared with other wild‐type Yarrowia lipolytica strains. Copyright © 2003 Society of Chemical Industry     

A water‐soluble supramolecular‐structured photoinitiator between methylated β‐cyclodextrin and 2,2‐dimethoxy‐2‐phenylacetophenone

A water‐soluble supramolecular‐structured photoinitiator (SSPI) was synthesized by supramolecular self‐assembling between methylated β‐cyclodextrin (MβCD) and hydrophobic 2,2‐dimethoxy‐2‐phenylacetophenone (DMPA). The structure of SSPI was characterized by X‐ray diffraction, FTIR, ¹H NMR, UV–vis, and fluorescence spectra. The results indicated that MβCD and DMPA had formed 1 : 1 inclusion complex in methanol solution. The binding constant (K) for the complex was 7.51 × 10²M^?1. SSPI could be dissolved in water easily and its water‐solubility was 15.3 g/100 mL. SSPI was the more efficient photoinitiator than DMPA for the photopolymerization of acrylamide (AM) in homogeneous aqueous system. The conversion for photopolymerization of trimethylolpropane triacrylate system initiated by SSPI was similar to that initiated by DMPA. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 2007     

Kinetics of Denaturation and Effects of Surfactants and Polyethylene Glycol on Soybean Esterase (Glycine max L) Stability

The objective of this work was to evaluate the kinetics and thermodynamics parameters and the effects of anionic, cationic and nonionic surfactants and polyethylene glycol on the activity and stability of a crude esterase extracted from soybeans (Glycine max L.). The activation energy for thermal inactivation was calculated from the Arrhenius plot was found to be 59.4 kJ mol^?1 and the ΔH* 56.82 kJ mol^?1 at 40 °C, which was the optimum temperature for enzyme activity. The ΔS* and ΔG* of the enzyme were found to be 61.67 kJ mol^?1 and 15.50 J mol^?1 K^?1, respectively, at the optimum temperature. The activity was only enhanced by the cationic surfactants cetyltrimethylammonium bromide and tetradecylmethylammonium bromide at a concentration of 3.0 mM. The anionic surfactant showed a positive effect on enzyme activity at the concentrations of 1.5 and 3.0 mM. Aqueous PEG (polyethylene glycols) solutions activated the esterase, and maximum activation (170 %) occurred with the addition of 6 kDa PEG. PEG with molecular weights of 0.4 and 10 kDa enhanced enzyme stability at 40 °C.     

Purification and characterization of a thermophilic chitinase produced by <Emphasis Type="Italic">Aeromonas</Emphasis> sp. DYU-Too7

An extracellular chitinase, produced by Aeromonas sp. DYU-Too7, was purified in the following procedures: ammonium sulfate precipitation, ultrafiltration, chromatographic separation of DEAE-sepharose CL-6B and sephacrylS-100HR. The resulting chitinase has a molecular mass of 36 kDa, an optimal reaction pH of 5.0, and an optimal reaction temperature of 70°C. It retains almost 100% activity in the pH range of 5.0–8.0. This chitinase has a high thermal tolerance and retained 90% of its activity at 50°C and 75% at 60°C. Enzyme activity was inhibited by Ba²⁺, Hg²⁺, Mg²⁺ and Ag⁺ cations, but was not substantially inhibited by the K⁺ cation nor the chelating agent EDTA. The K _m and V _mα , using colloidal chitin as a substrate, are 6.3 g/L and 18.69 μmol/min/mg-protein, respectively. The 36 kDa chitinase of Aeromonas sp. DYU-Too7 is an exo-type enzyme, because chitobiose was the main hydrolysate in hydrolysis of colloidal chitin.     

Characterization of cellulase and xylanase activities of Clostridium celerecrescens

The production of cellulases and xylanase by Clostridium celerecrescens, a new anaerobic mesophilic cellulolytic bacterium, was studied using various substrates (cellobiose, xylan and cellulose Whatman CF-11). While both cellulase (β-1,4-D-glucan glucanohydrolase) and xylanase (β-1,4-xylan xylanohydrolase) were produced on cellulose, only the latter was produced when xylan was used as the sole carbon source. A weak p-nitrophenyl-β-D cellobiohydrolase activity was detected in the extracellular filtrates when using cellulose as a substrate. Otherwise, β-glucosidase (p-nitrophenyl-β-D-glucopyranosidase) was always found to be associated with the bacteria and reached its maximum levels of growth on cellobiose. In all cases, enzyme production showed a cell growth associated profile. Activities of these enzymes had their optimal values within the ranges of temperature and pH reported for the corresponding enzymes from similar anaerobic mesophilic microorganisms, although a relatively high optimum temperature, 55°C, was found for xylanase. All enzymes showed a 90% reduction of half-life time for each 8°C increment of temperature. A 50% inhibition of xylanase and β-cellobiohydrolase activity was observed, through a competitive mechanism, by xylose (0.677 mmol dm^?3) and cellobiose (28 mmol dm^?3) respectively.     

Study on microbial protein and the mechanism of solid-state fermentation with periodical dynamic changes of air

The effect of different extraction and purification conditions on the microbial protein obtained from solid-state fermentation (SSF) and the effect of periodical dynamic changes of air on protein have been studied. The mechanism of solid-state fermentation with periodical dynamic changes of air is also discussed. Compared with static solid-state fermentation, periodical dynamic changes of air afford a higher protein mass; from 1 g of the fermentation microbe, 5.3 mg of the intracellular protein (an increase of 34.63%) and 9.09 mg of the extracellular protein (an increase of 17.8%) were obtained on the sixth day of fermentation. The filter paper activity and carboxymethyl cellulose activity (FPA and CMCase) of the extracellular protein are 1.739 μmol/s and 109.592 μmol/s, respectively, which represent increases of 60.1% and 21.2% over the corresponding values for static solid-state fermentation. The FPA and CMCase of the intracellular protein are 0.245 μmol/s and 6.392 μmol/s, respectively, which represent decreases of 22.2% and 38.7% over the corresponding values for static solid-state fermentation. The enzyme activity of the microbial extracellular protein in solid-state fermentation with periodical dynamic changes of air on the fifth day is nearly equal to that on the sixth day without periodical dynamic changes of air, so the period of fermentation can be shortened. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) experiments suggest that pulsating air pressure stimulation leads to a decrease in the amount of protein component with molecule mass of about 80400Da, and an increase in the amount of protein component with molecule mass of about 28520Da. Translated from Journal of Beijing University of Chemical Technology, 2006, 33(3): 42–46 [译自: 北京化工大学学报]