Neutral Enzyme Research Articles

Study on the effect and mechanism of ultrasonic-assisted enzymolysis on antioxidant peptide activity in walnuts

Walnut meal is a large quantity and high-quality resource with great exploitation value. Ultrasonic-assisted enzymolysis (UAE) was utilized in the preparation of peptides from walnut meal protein. Results indicated that by optimizing the UAE process with neutral protease, an ultrasound power of 180 W, a 4.3 h duration and an enzyme dosage of 10 KU/g, the walnut peptides exhibited the most potent antioxidant activity. In comparison to the control group, the WPI treated with ultrasound and neutral enzymes in combination (UNWPI) demonstrated a significant enhancement in their DPPH, ABTS, and ·OH scavenging capabilities, with increases of 234.23 %, 240.22 %, and 69.52 %, respectively. By analyzing the structure of walnut antioxidant peptides with or without ultrasound, it was observed that the underlying mechanism for the increased antioxidant activity was that UAE not only formed more small peptides, but also produced more peptides with hydrophobic amino acids at their terminal ends. Subsequently, six peptides were identified and screened from UNWPI, namely IFW, IIPF, IVAF, IIFY, ILAFF, and IFIP, which exhibited high antioxidant activity and could bind to Keap1 protein through hydrogen bonding, π-alkyl interactions, and π-π stacking interactions. The research results provided theoretical basis and technical support for the preparation of walnut antioxidant peptides and the high-value utilization of walnut meal.

Biochemical Characterization and Application of Zearalenone Lactone Hydrolase Fused with a Multifunctional Short Peptide.

Zearalenone (ZEN) is an estrogenic mycotoxin causing reproductive toxicity in livestock. Currently, lactone hydrolases are used in the enzymatic degradation of ZEN. However, most lactone hydrolases suffer from low degradation efficiency and poor thermal stability. ZHD518, as a documented neutral enzyme for ZEN degradation, exhibits high enzymatic activity under neutral conditions. In this study, a multifunctional peptide S1v1-(AEAEAHAH)2 was fused to the N-terminus of ZHD518. Compared with the wild-type enzyme, the peptide fusion significantly enhanced protein expression by 1.28 times, enzyme activity by 9.27 times, thermal stability by 37.08 times after incubation at 45 °C for 10 min and enzyme stability during long-term storage. Moreover, ZEN concentrations in corn bran, corn germ meal, and corn gluten powder decreased from 5.29 ± 0.04, 5.31 ± 0.03, and 5.30 ± 0.01 μg/g to 0.48 ± 0.05, 0.48 ± 0.06, and 0.21 ± 0.04 μg/g, respectively, following a 60 min treatment with S1v1-GS-ZHD518, resulting in degradation rates of 90.98, 91.00, and 95.32%, respectively. In conclusion, the properties of S1v1-GS-ZHD518, such as its efficient degradability, high temperature resistance and storage resistance, offer the possibility of its application in food or feed.

Biochemical Characterization of a Marine Pseudoalteromonas citrea-Derived Fatty Acyl-AMP Ligase That Exhibits N-Acyl Amino Acid Synthetic Activity.

Activation of fatty acids as acyl-adenylates by fatty acid-AMP ligase (FAAL) is a well-established process contributing to the formation of various functional natural products. Enzymatic characterization of FAALs is pivotal for unraveling both the catalytic mechanism and its role in specific biosynthetic pathways. In this study, we recombinantly expressed and characterized a novel FAAL derived from marine Pseudoalteromonas citrea (PcFAAL). PcFAAL was a cold-adapted neutral enzyme, demonstrating optimal activity at 30°C and pH 7.5. Notably, its specific activity relied on the presence of Mg2+; however, higher concentrations exceeding 10mM resulted in inhibition of enzyme activity. Various organic solvents, especially water-immiscible organic solvents, demonstrated an activating effect on the activity of PcFAAL on various fatty acids. The specific activity exhibited a remarkable 50-fold increase under 4% (v/v) n-hexane compared to the aqueous system. PcFAAL displayed a broad spectrum of fatty acid substrate selectivity, with the highest specific activity for octanoic acid (C8:0), and the catalytic efficiency (kcat/Km) for octanoic acid was determined to be 1.8nM-1·min-1. Furthermore, the enzyme demonstrated biocatalytic promiscuity in producing a class of N-acyl amino acid natural products, as verified by LC-ESI MS. Results indicated that the PcFAAL exhibits promiscuity towards 10 different kinds of amino acids and further demonstrated their potential value in the biosynthesis of corresponding functional N-acyl amino acids.

Fabrication and evaluation of some characteristics of the skin's dermis gel direction to application in regenerative medicine

Objective:The tight connection between the cell-extracellular matrix (ECM - Extracellular matrix) is the foundation of tissue development and repair. The extracellular framework simulates the biological properties of organism-based ECM. The ECM of the skin’s dermis is a natural material, containing ingredients ideal for creating products that help tissue heal. This study focuses on the production of gel products from human skin ECM. The skin ECM is gelled by dissolving in a pepsin enzyme solution.Method: 3D gel formation process using neutral enzyme solution and incubation under physiological conditions. Gel samples were evaluated for speed and formability. Additionally, the dermal gel preparation was evaluated for its effects on the survival and proliferation of human fibroblasts in vitro by toxicity testing and proliferation testing, for 10 days. Results: Direct and extract toxicity assessments showed that the ECM gel regimen was safe for fibroblast survival. At the same time, fibroblasts had proliferation properties after 10 days in culture conditions with gel extract.Conclusion: These results showed that the ECM gel product regimen has the ability to positively impact the biological properties of cells and has the potential for application in the field of regenerative medicine.

Progress in the Conversion of Ginsenoside Rb1 into Minor Ginsenosides Using β-Glucosidases.

In recent years, minor ginsenosides have received increasing attention due to their outstanding biological activities, yet they are of extremely low content in wild ginseng. Ginsenoside Rb1, which accounts for 20% of the total ginsenosides, is commonly used as a precursor to produce minor ginsenosides via β-glucosidases. To date, many research groups have used different approaches to obtain β-glucosidases that can hydrolyze ginsenoside Rb1. This paper provides a compilation and analysis of relevant literature published mainly in the last decade, focusing on enzymatic hydrolysis pathways, enzymatic characteristics and molecular mechanisms of ginsenoside Rb1 hydrolysis by β-glucosidases. Based on this, it can be concluded that: (1) The β-glucosidases that convert ginsenoside Rb1 are mainly derived from bacteria and fungi and are classified as glycoside hydrolase (GH) families 1 and 3, which hydrolyze ginsenoside Rb1 mainly through the six pathways. (2) Almost all of these β-glucosidases are acidic and neutral enzymes with molecular masses ranging from 44-230 kDa. Furthermore, the different enzymes vary widely in terms of their optimal temperature, degradation products and kinetics. (3) In contrast to the GH1 β-glucosidases, the GH3 β-glucosidases that convert Rb1 show close sequence-function relationships. Mutations affecting the substrate binding site might alter the catalytic efficiency of enzymes and yield different prosapogenins. Further studies should focus on elucidating molecular mechanisms and improving overall performances of β-glucosidases for better application in food and pharmaceutical industries.

The Impact of Neutral Enzyme on Jute-Cotton Blended Denim Fabric

Garments washing is the process of altering the aesthetic look, luxury, fashion, and comfortability of a garment after it has been manufactured. Most garments nowadays go through many washing processes before being sold in retail outlets. As denim is one of the most adaptable fabrics available today, the aesthetics may be reinvented in an infinite number of ways through garments washing techniques. Because of the rough and stiff nature of jute fiber, it is not currently used as a popular material for garments manufacturing. A sustainable option may be possible by using jute and cotton blended fabric in manufacturing. Moreover, enzyme is a naturally occurring product. Therefore, the product is environmentally friendly. In a washing bath, the enzyme functions as a catalyst to hydrolyze fiber, soften the fiber surface, and decrease pilling. The experiment was conducted on a 70 percent cotton, 30 percent jute blended fabric with a weave construction of 2/1 twill. The goal of this study is to observe how an industrial enzyme wash affects a jutecotton blended denim fabric. For this investigation, neutral enzyme was used. Finally, the raw and washed samples was compared in terms of physical parameters such as EPI, PPI, GSM, warp strength, and weft strength. This research may unlock a way for enzymatic treatment on jute cotton blended denim fabric. GUB JOURNAL OF SCIENCE AND ENGINEERING, Vol 8, Dec 2021 P 24-28

Study of the effect of enzymatic washing parameters on the bagging properties of denim fabric with Taguchi method

AbstractThis study aims to evaluate the effect of neutral enzyme washing on the bagging behavior of denim garments and optimize the conditions of this treatment using the Taguchi design. Six types of fabrics were selected with the same weaves and different characteristics. These samples were washed with a neutral cellulase enzyme under different conditions such as enzyme concentration (0.3%, 2%, and 4%), temperature (30, 45, and 65°C), time (15, 45, 75 min), and material to liquor ratio (1:5, 1:8, and 1:10). To evaluate the effect of enzymatic washing on the bagging behavior, we investigated the residual bagging height, the bagging fatigue percentage, the bagging resistance, and the bagging recovery percentage. Thanks to the main effects plots the influencing factors were obtained and using the signal‐to‐noise ratio diagram, the optimum conditions were determined. The obtained results showed that these bagging properties were affected by the washing conditions and that the most important factors are the characteristics of the samples such as the percentage of elastane and polyester fibers, weight and weft density, and enzyme concentration. The process parameters were optimized and the optimized washing conditions for the best value were 0.3% enzyme concentration at 45°C for 75 min, in material to liquor ratio of 1:8. Concerning the factor of the sample, the results show that the increase of the percentage of elastane or polyester fibers in the composition of denim fabric decreases the problem of deformation and bagging. Therefore it is recommended to add a percentage of these fibers in the construction of the denim to improve the bagging recovery. Thus, the increase in the weft density minimizes that bagging phenomen appears.

Enzymatic Hydrolysis of Broken Rice Protein: Antioxidant Activities by Chemical and Cellular Antioxidant Methods.

Excessive reactive oxygen species (ROS) is an important cause of aging, and supplementing antioxidants through diet is one of the important ways to delay aging. Some studies have confirmed that rice protease hydrolysate has antioxidant activity, but was rarely been investigated on cells. Thus, commercial enzymes, alkaline enzyme, neutral enzyme, pepsin, chymotrypsin, and trypsin were selected to hydrolyze broken rice protein (BRP) to obtain the corresponding hydrolysates, which were A-broken rice protein hydrolysate (BRPH), N-BRPH, P-BRPH, C-BRPH, and T-BRPH, respectively. Then the antioxidant properties of BRPHs were evaluated by different chemical and cellular antioxidation. Molecular weight, peptide length distribution, and amino acid sequence were detected to insight into the antioxidant properties. Among BRPHs, the A-BRPH displayed the strongest hydroxyl radical scavenging activity (IC50 = 1.159 mg/ml) and metal ion-chelating activities (IC50 = 0.391 mg/ml). Furthermore, cellular antioxidation confirmed that A-BRPH significantly increased cell viability and inhibited the intracellular ROS release in both aging cells and cell-aging processes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results revealed that peptides with molecular weight <14.5 KDa were produced by enzymatic hydrolysis. Additionally, A-BRPH rich in low molecular weight (<3 kDa) and short-length peptides with some specific amino acids, such as aromatic and hydrophobic amino acids, contributes to the antioxidant properties. This study provided theoretical to the utilization of broken rice and confirmed that A-BRPH could be used in new anti-aging food and health products for human consumption.

A thermostable extracellular α-amylase from Aspergillus flavus S2-OY: Purification, characterisation and application in raw starch hydrolysis

An amylolytic fungus was isolated locally from the soil of a cassava waste dumpsite. It was identified as Aspergillus flavus S2-OY using 18S rRNA gene sequencing with gene bank accession number MZ267053.1. α-Amylase was produced from the fungus through solid substrate fermentation of potato (Solanum tuberosum L.) peel, a cheap and renewable substrate. The fungal enzyme was purified 13.41-fold by ammonium sulphate and ion exchange chromatography (CM Sephadex C-50), with final yield of 13.41%. Apparent molecular weight of 55.0 kDa was obtained for the purified enzyme, using SDS-PAGE. Optima pH (6.0) and temperature (70 °C) of α-amylase activity were determined for the enzyme which also showed thermal stability up to 70 °C, with 80% residual activity after 120 min. The enzyme exhibited a wide range of pH stability (4.0 to 7.0). Kinetic parameters Km and Vmax values obtained with soluble starch were 158 µg/ml/min and 1.51 mg/ml, respectively. The enzyme was slightly inhibited by Mg2+. Mn2+, Zn2+, Na+, K+, Hg2+ and the chelating agent, EDTA, at 5.0 mM concentration. However, it was activated by Sn2+. The enzyme hydrolysed gelatinised and ungelatinized raw yam, corn, rice and millet starches strongly but cassava starch moderately. These results revealed that cost-effective α-amylase production from Aspergillus flavus S2-OY could be realised with the use of potato peel as a cheap substrate, under solid substrate fermentation condition. Also, the fungal α-amylase is a raw starch-hydrolysing, acid to neutral and thermostable enzyme with potential for broad biotechnological applications.

Antioxidant and functional properties of cowhide collagen peptides.

In the present study, antioxidant activities and functional properties of cowhide collagen antioxidant peptides (CCAPs) with different molecular weight (MW) were investigated. The optimum preparation conditions of CCAPs were hydrolysis time of 1.53 hr, temperature of 54.9 °C, pH 7.38, and neutral enzyme to trypsin ratio of 0.048g: 0.016g according to single factor test and response surface methodology (RSM). Three fractions (CCAP-I, CCAP-II, and CCAP-III) were obtained by ultrafiltration and lyophilization. Antioxidant activities revealed that CCAP-III had high reducing power activity (0.323± 0.035) and scavenging effect on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals (64.30± 5.99%), 2,2-azino-bis-(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) radicals (75.25± 3.14%), and hydroxyl radicals (68.26± 6.74%) compared to the other fractions. In addition, LC-MS/MS analysis showed that Ala-Gly-Glu-Arg, Gly-Ile-Ala-Gly-Glu-Arg, Gly-Pro-Ala-Gly-Pro-Ala-Gly-Pro-Arg, Gly-Val-Val-Gly-Pro-Glu-Gly-Ala-Arg and Gly-Phe-Ser-Gly-Leu-Asp-Gly-Ala-Lys were the major peptides of CCAP-III. CCAP-III showed good hygroscopicity (HYG), water holding capacity (WHC), and oil holding capacity (OHC) when compared with CCAP-I and CCAP-II. However, CCAP-II has great emulsifying properties, and CCAP-I has excellent foaming properties. Therefore, CCAPs can be used as a promising source of functional peptides with antioxidant properties. PRACTICAL APPLICATION: This study demonstrated the peptides of cowhide collagen has superior antioxidant and functional properties. This study provided a scientific basis for the preparation of antioxidant peptides from cowhide collagen.

Characterization of a novel Cu-containing dissimilatory nitrite reductase from the haloarchaeon Halorussus sp. YCN54.

Dissimilatory nitrite reductase, a key enzyme in the denitrification pathway, catalyzes the reduction of nitrite to NO. Bioinformatic analysis showed that the genome of a novel nitrite-degrading haloarchaeon Halorussus sp. YCN54 possessed a gene encoding the Cu-containing dissimilatory nitrite reductase (NirKHrs). NirKHrs was heterologously expressed and purified. Protein sequencing indicated that two isoforms of NirKHrs monomer were produced intracellularly. UV-vis spectrum of the purified NirKHrs showed that it belonged to the blue NirK group. NirKHrs showed optimum activity at 4.5M NaCl, 55 ℃ and pH 7.0, representing a halophilic, slightly thermophilic and neutral enzyme. It exhibited high stability at 30-50 ℃. NirKHrs activity was strongly inhibited by the copper chelating agent due to removal of copper. NirKHrs activity was activated by Mn2+ and Sr2+. It displayed good tolerance to some high polarity organic solvents and nonionic surfactants, such as glycerol, DMSO, DMF and tween-20. Na2S2O4 was an effective electron donor to NirKHrs. The Km and Vmax values of purified NirKHrs for nitrite were 3.2mM and 477.2 U/mg, respectively, indicating its high activity. These results indicated that NirKHrs may have potential applications for nitrite degradation in high-salt industries, such as salted food and saline wastewater treatment.

Effect of protease and phytase on the physiological state of alcoholic yeast in cultivation

Saccharomyces cerevisiae yeast is used in the production of ethyl alcohol. The main requirements for yeast used in the production of ethyl alcohol from starch-containing raw materials: alcohol yeast used in the processing of starchy raw materials must have high fermentation activity; complete fermentability of sugars, resistance to metabolic products, resistance to the development of extraneous microflora. Proteolytic enzyme preparation Prolive BS Liquid was used as a source of protease. Kingphos enzyme preparation was used as a source of phytase. The effect of the enzyme preparations of the neutral protease Prolive BS Liquid and Phytase Kingphos on the fermentation activity of alcohol yeast Saccharomyces cerevisiae race XII was studied. The maximum fermentation activity is possessed by yeast cultivated on the wort, obtained using protease and phytase. The duration of the exponential growth phase in the experiment is 14–16 hours, in the control -18–20 hours. The exponential phase is described by the Mono equation. Compared to the yeast in the control, the yeast in the experiment multiplies more intensively, and by 14–16 hours of growth, about 170 million yeast cells accumulate in the culture medium, and the yeast in the control-about 95 million yeast cells by 18–20 h of growth. The specific growth rate was maximum in the logarithmic phase and amounted to 0.35 million cells / cm3 • h in the experimental samples and 0.25 million cells / cm3 • h in the control. It was found that the maximum accumulation of yeast cells was observed when the neutral enzyme Prolive BS Liquid was added to the wort with a dosage of 0.2 units. PS/g of starch and enzyme preparation Phytase Kingfos with a dosage of 0.5 units. FS/g of starch, the yeast cell content in mature yeast reached 170 million cells / cm3 by 16-18 hours of cultivation, the yeast has a high fermentation activity.

(Invited) Investigating the Hydrogenase Mechanism By Protein Film Electrochemistry

With turnover frequencies up to 106 s-1, hydrogenases are the fastest molecular hydrogen catalyst. They are classified according to the metal content of their active, distinguishing the NiFe and FeFe as the two major classes. The two classes display an outstanding organometallic cofactor, with cyanido- and carbonyl- groups coordinating the metals. Their mechanism includes the stabilization of a metal-hydride state as the most reduced intermediate of the catalytic cycle. Hydrogenase (Hase) have driven mechanistic studies for decades, including extensive works by protein film electrochemistry targetting aspects as diverse as intramolecular diffusion of gases, long distance electron transfer, or susceptibility to inhibitors. This communication will focus on two other questions, distinguishing steady state functioning and in-activation processes (see figure). The catlaytic bias is the ability of an enzyme to turnover faster one direction than another. Amongst hydrogenases, the catalytic bias spans full range, from extremely biased to essentially neutral enzymes. Several works have brought clues to understand the molecular basis of the catalytic bias for both Hase classes, pointing out to the intramolecular electron transfer chain as the main effector. Our recent results on the study of the hydrogen dependent CO2 reductase reveal the role of the affinity for hydrogen in the bias. The susceptibility to overpotential corresponds to in-activation processes triggered by redox conditions that are far from equilibrium, and plausibly encountered in the life cycle of bioelectrochemical devices. One particular phenomenon is the reductive inactivation of FeFe-Hases, undergoing both in solution and on the electrode, that remains largely to understand. Our works on the enzyme HydA1 from microalgae suggest that this inactivation can be reversed under maintained reductive conditions, unlike discussed in the literature. Figure 1

Enzymatic identification and functional sites study of a novel cold-active cellulase (MkCel5) from Microbacterium kitamiensea

A strain of Microbacterium kitamiense capable of decomposing cellulose was obtained from the soil of Shigatse region in Tibet. Low temperature and cellulase gene MkCel5 was cloned and sequenced. The size of the open-reading frame was 1449 bp; it was predicted to encode a polypeptide with 482 residues with a molecular weight of 51.38 kDa. Sequence analysis indicated that it belongs to the glycoside hydrolase family 5 (GH5), which has high homology with reported endoglucanases. The predicted protein structure had a typical carbohydrate-binding domain and a catalytic domain of cellulase. Site mutagenesis experiments showed that the two glutamates at positions 312 and 404 were crucial in the catalytic process. MkCel5 was identified as a cold-adaptive and neutral enzyme with a maximum activity at 25 °C and pH 7.0. These properties suggested that MkCel5 has potential for application in the cellulose processing industry requiring low temperature.

Effect of Cellulase Enzymes and Swelling Agents on Colour Strength and Colourfastness of Handloom Cotton Fabric Dyed with Butea Frondosa

Cellulases and swelling agents are known to be effective in improving the colour strength of cotton. Nowadays, handloom fabrics, such as khadi cotton, are much more preferred due to the development of innovative designs with their use and their comfort in wearing. Also, due to increased environmental awareness, the use of natural dyes are much more preferred in the dyeing of handloom fabrics. However, khadi cotton has some major shortcomings, such as less dyeability. The present study is carried out by keeping in mind that the pretreatment of khadi cotton with cellulases, swelling agents and a combination of cellulases and swelling agents before dyeing improves the colour strength properties. Khadi cotton samples are treated with optimized conditions of the enzymes and swelling agents. The optimum pH, concentration, treatment time and temperature selected for treatment of the samples with acid cellulase enzymes are 5.5, 1.5% (owf), 45 minutes and 50°C, respectively, whereas in the case of neutral cellulase enzymes, 7.5, 2.0% (owf), 70 minutes and 70°C, respectively. The optimum concentration, treatment time and temperature selected for the treatment of the samples with sodium hydroxide, ethylenediamine and zinc chloride are 20% w/v, 60 minutes and 60°C; 80% w/v, 60 minutes and 70°C, and 80% w/v, 60 minutes and 70°C respectively. Butea frondosa dye (5 g) extracted for 75 minutes provides the best results on khadi cotton when dyeing is carried out for 90 minutes. It is observed that out of the various concentrations of mordants used with the Butea frondosa dye, the best shades of colour are obtained by using 0.04 g of alum, 0.01 g of copper sulphate, and 5 g of Babool bark. In terms of optimizing the mordanting, the best results are obtained with Butea frondosa dye when the samples are simultaneously mordanted and dyed with alum, Babool bark and alum. Pre-mordanting is selected for the copper sulphate. It is found that for all the enzyme treated (acid and neutral cellulase) as well as swelling agent treated (sodium hydroxide, ethylenediamine and zinc chloride) samples, the colour strength and colourfastness increase in comparison to the untreated samples.

EFFECT OF CELLULASE ENZYMES AND SWELLING AGENTS ON COLOUR STRENGTH (K/S) AND COLOURFASTNESS PROPERTY OF HANDLOOM COTTON FABRIC DYED WITH BUTEA FRONDOSA DYE

Cellulase and swelling agents are known to be effective in improving the colour strength of cotton. Nowadays, the handloom fabrics are much preferred due to development of innovative designs and their comfort in wearing Due to increased environmental awareness the use of natural dyes are much preferred in dyeing of handloom fabrics. But, khadi cotton has some major shortcomings like less dyeability. Keeping in view that the pretreatment of khadi cotton with cellulases, swelling agents and combination of cellulases and swelling agents before dyeing improves the colour strength properties, the present study was planned. The khadi cotton samples were treated with optimized conditions of enzymes and swelling agents. The optimum pH, concentration, treatment time and temperature selected for the acid cellulase enzyme treatment were 5.5, 1.5% (owf), 45 minutes and 50°C, respectively whereas in case of neutral cellulase enzyme, it was 7.5, 2.0% (owf), 70 minutes and 70°C respectively. The optimum concentration, treatment time and temperature selected for sodium hydroxide treatment were 20% w/v, 60 minutes and 60°C, respectively. In case of ethylenediamine, 80% w/v, 60 minutes and 70°C were selected as optimum concentration, treatment time and temperature, respectively. In case of zinc chloride treatment, the optimum concentration, treatment time and temperature were selected as 80% w/v, 60 minutes and 70°C. 5 g Butea frondosa dye extracted for 75 minutes gave best results on khadi cotton when dyeing was carried out for 90 minutes It was observed that out of various concentrations of mordants used with Butea frondosa dye, best shades of colour were obtained by using 0.04 g of Alum, 0.01 g of Copper sulphate, 5 g each of Baboolbark.On optimizing the method of mordanting, best results were obtained with Butea frondosa dye when samples were simultaneous mordanted and dyed with Alum, Babool bark and Alum. Pre-mordanting was selected for Copper sulphate. . It was found that for all the enzyme treated (acid and neutral cellulase) as well as swelling agents treated (Sodium hydroxide, Ethylenediamine and Zinc chloride) samples, the colour strength and colourfastness was increased in comparison to the untreated samples.

Impact of Washing Stretchy Denim using Neutral and Acid Enzymes and Subsequent Softening Treatment on Physical, Mechanical and Sewing Properties

Comfort, versatility and durability continue to be stretchy denim’s biggest assets, therefore it is known for its use in the manufacture of jeans; however, it can be used to make several apparels, depending on its mass. Even designers turn to it for inspiration occasionally. The main factors affecting consumers when selecting garments are aesthetic appearance and fashion. Denim garments are subjected in industrial washing to obtain specific appearance and handle. The washing and finishing processes are utilized for the purpose of fashion and different recipes are applied for different effects which are quite significant for marketing. Washing is a novel process to impart worn-out look, to modify the appearance and to improve the comfort ability of apparel. Enzyme wash is used to fade the color of denim as well as it has an effect on the physical, mechanical and sewing properties of the denim also. By this paper washing of tested denim with enzyme supplemented with softening were applied to preserve the quality and to create more distress worn-look fashionable denim. It presents the impact of enzyme wash type and subsequent softening treatment on stretchy denim dyed with indigo dye. Two different masses (380 & 320 gm/m2) of stretchy denim were sewed separately with lockstitch 301 by using two types of seams (lapped seam and flat felled seam) and washed using neutral and acid enzyme wash and then softened using silicone softener. The physical, mechanical and sewing properties were tested before washing, after washing and after softening. The properties that were analyzed include fabric mass, thickness, stiffness, tensile strength, seam strength, seam pucker and seam appearance. Neutral enzyme washed, acid enzyme washed and softened tested denim exhibit a clear difference in the tested properties than the unwashed denim. It’s concluded that neutral enzyme wash supplemented with softening gives it the best value compared to all the tested denim washed properties

Etiology screening role of transrectal ultrasonography in male obstructive azoospermia infertility

Objective To study etiology screening role of transrectal ultrasonography in male obstructive azoospermia infertility. Methods The clinical data of 328 cases who suspected of being obstructed sperm disease were retrospectively analyzed. TRUS detection was conducted, at the same time, the sperm amount, sperm and semen p H, pure berries quantitative, neutral sugar alpha glycosidase enzymes quantitative, elastic hard protease were tested. Results In 328 cases with male obstructed no sperm,by TRUS detection results,216 cases(65.8%)could find the causes, ejaculatory duct expansion, seminal vesicle gland lesions, prostate midline cyst were the top three causes respectively; 112 patients(34.2%) had no obvious abnormal ultrasonic testing. Sperm was not seen in semen of obstructive azoospermia patients and semen p H<7,pure berries sugar quantitative and quantitative value neutral alpha glycosidase enzymes were very low,hard elastic protease was low. Conclusion The main causes of obstructive azoospermia were ejaculatory duct expansion, seminal vesicle gland lesions, prostate midline cyst, sperm TRUS detection used for diagnosis of high sensitivity, and easy to operate, noninvasive, and combined with seminal plasma biochemical examination, the diagnostic effect is much better. Key words: Infertility; Azoospermia; Ultrasonography

Optimization of Enzymatic Preparation Anti-Oxidation Peptides from Pigskin Gelatin

We screen the neutral protease enzyme used to hydrolysis, through scavenging effect of six enzymatic hydrolysate on O2-•. The hydrolysis of condition was optimized by response surface methodology on the basis of single factor experiments. The optimized condition was as follows: hydrolysis temperature 43.65 °C, pH 6.25, time 3.29h, and content of enzyme 3.0% (W/W). Under this condition, the O2-•scavenging rate of the hydrolyzate was up to 72.01%. The experimental results were 71.15%.

Biochemical properties of an extracellular trehalase from Malbranchea pulchella var. Sulfurea

The thermophilic fungus Malbranchea pulchella var. sulfurea produced good amounts of extracellular trehalase activity when grown for long periods on starch, maltose or glucose as the main carbon source. Studies with young cultures suggested that the main role of the extracellular acid trehalase is utilizing trehalose as a carbon source. The specific activity of the purified enzyme in the presence of manganese (680 U/mg protein) was comparable to that of other thermophilic fungi enzymes, but many times higher than the values reported for trehalases from other microbial sources. The apparent molecular mass of the native enzyme was estimated to be 104 kDa by gel filtration and 52 kDa by SDS-PAGE, suggesting that the enzyme was composed by two subunits. The carbohydrate content of the purified enzyme was estimated to be 19 % and the pi was 3.5. The optimum pH and temperature were 5.0-5.5 and 55° C, respectively. The purified enzyme was stimulated by manganese and inhibited by calcium ions, and insensitive to ATP and ADP, and 1 mM silver ions. The apparent K(M) values for trehalose hydrolysis by the purified enzyme in the absence and presence of manganese chloride were 2.70 ± 0.29 and 2.58 ± 0.13 mM, respectively. Manganese ions affected only the apparent V(max), increasing the catalytic efficiency value by 9.2-fold. The results reported herein indicate that Malbranchea pulchella produces a trehalase with mixed biochemical properties, different from the conventional acid and neutral enzymes and also from trehalases from other thermophilic fungi.