Commercial Enzyme Research Articles

The effects of exogenous dietary enzyme on the growth of giant gourami (Osphronemus goramy)

The commercial complex enzymes containing protease, lipase, and amylase are potentially used for exogenous dietary enzymes on giant gourami (Osphronemus goramy) as growth booster indirectly. The aims of this study are to know the effect of the exogenous dietary enzymes on giant gourami growth and to obtain the optimal dose of this enzyme. The research was conducted on January-April 2019 in the aquaculture facility and laboratory of Universitas Airlangga, Banyuwangi Campus. A total of 200 giant gourami juveniles (5.5 ± 0.5 cm in total length) were obtained from the fish hatchery centre of Kabat, Banyuwangi, Indonesia. All fish was divided into 20 glass aquaria (40 x 30 x 30 cm3) and reared for 40 days with exogenous dietary enzyme treatments in the diets following: control (no enzyme), 2.5 ml/100 g feed, 5.0 ml/100 g feed, 7.5 ml/100 g feed, 10.0 ml/100 g feed (4 replication in each treatment). The parameters were observed in this study are daily growth rate (DGR), specific growth rate (SGR), and feed conversion ratio (FCR). The exogenous dietary enzyme significantly (P < 0.05) effect to all parameters, the best result was showed in 10.0 ml/100 g feed treatment as optimum applicate dose.

Low-cost homemade cocktails for enzymatic conversion of sugarcane and cassava bagasses

ABSTRACT The agricultural industries generate lignocellulosic wastes that can be modified by fungi to generate high value-added products. This work aimed to analyze the efficiency and the cost-effectiveness of the bioconversion of sugarcane and cassava bagasses using low-cost homemade enzymatic cocktails from Aspergillus niger LBM 134. Both bagasses were pretreated with a soft alkaline solution without any loss of polysaccharides. After the hydrolysis, a 28% of conversion to glucose and 42% to xylose were reached in the hydrolysis of sugarcane bagasse while an 80% of saccharification yield, in the hydrolysis of cassava bagasse using the homemade enzymes. Furthermore, a more disorganised surface and no starch granules were observed in the sugarcane and cassava bagasses, respectively. The bioethanol yield from sugarcane and casava bagasses was predicted to be 4.16 mg mL−1 and 2.57 mg mL−1, respectively. A comparison of the cost of the homemade and the commercial enzymes was carried out. Similar hydrolysis percentages were achieved employing any enzyme; however, it was 1000–2000 times less expensive using the homemade cocktails than using the commercial enzymes. Therefore, the cost of obtaining glucose from bagasses was most expensive when applying the commercial enzymes. Moreover, the hydrolysis of the cassava bagasse was most efficient with the homemade cocktails. The importance and novelty of this work lie in the similar performance and the lower cost of the homemade cocktails from the fungus A. niger LBM 134 compared with the commercial enzymes on the hydrolysis of the sugarcane and cassava bagasses.

Stain-removal potential of keratinase produced by Bacillus species isolated from feather waste

The aim of this study was to determine the potential of keratinases produced by Bacillus species isolated from poultry feather waste in stain removal. The effect of individual and combined detergent components on the keratinase activity of EZYKer-51, EZYKer-50 and EZYKer-53 enzymes produced from keratin degrading Bacillus licheniformis-K51, Bacillus subtilis-K50 and Bacillus sp.-K53 respectively was determined. EZYKer-51, which had the least enzyme inhibition in the presence of the detergent component was used as the enzyme component in detergent formulation and a wash performance analysis test was carried out to compare the stain removal capacity of DETKER 51 (detergent formulated using EZYKer-51 as enzyme component) with a commercial enzyme. Data were analyzed using descriptive statistics. EZYKer-51 showed a significantly higher (P≤0.05) enzyme activity in the presence of the detergent ingredients for both the crude and purified forms of the enzyme compared with the activities of EZYKer-50 and EZYKer-53, except for the activities (8.02±1.32 U/mL and 7.94±0.63 U/mL) in the presence of 15% LAS and 5% liquid paraffin. Visually, better stain removal was achieved when endogenous protease of commercial enzyme was replaced with EZYKer-51. These results suggest that keratinase EZYKer-51 may be a useful alternative for applications in detergent formulation.

Two-step pretreatment for improving enzymatic hydrolysis of spent coffee grounds

Spent coffee ground has attracted increasing attentions since it contains many useful components such as polysaccharides, protein, lipid and bioactive compounds. The aim of this research is to enhance the enzymatic hydrolysis to release important sugars in spent coffee ground using different pretreatment methods. Spent coffee grounds were pretreated by alkali pretreatment, organosolv pretreatment and the combined process. The pretreated material was hydrolyzed by different commercial enzymes including Cellulast, Pectinex, Ultraflomax and Viscozyme. Monosaccharides, total phenolic content and antioxidant activity in the hydrolysate were measured and evaluated. The use of Viscozyme achieved the highest reducing sugar yield and showed the significant difference from other enzymes. Alkali and organosolv pretreatment demonstrated to improve the production of sugars. The alkali pretreatment followed by organosolv treatment effectively removed lignin, resulting in only 14% lignin in the pretreated sample. The maximum reducing sugar concentration reached 6120 mg/L through two-step pretreatment and subsequent enzymatic hydrolysis, corresponding to a yield of 161 mg sugar/g substrate. The spent coffee ground hydrolysate contained 2917 mg/L mannose, 1633 mg/L glucose and 957 mg/L galactose. Phenolic compounds were observed to be released during the enzymatic hydrolysis, giving a total phenolic content of 174.4 mg GAE/L and the SCG hydrolysate also showed an antioxidant capacity equivalent to 263.2 mg/L ascorbic acid after 120 h hydrolysis. This study demonstrated a scalable two-step pretreatment process to obtain important sugars including mannose, glucose, and galactose along with phenolic compounds for further industrial uses.

Evaluation of substance P and bradykinin levels in nasal secretions of patients with nasal polyposis with and without sensitivity to non-steroidal anti-inflammatory drugs.

ObjectiveThe role of neurogenic inflammation in pathogenesis of chronic rhinitis is well known. However, very little is known about its importance in pathogenesis of nasal polyposis (NP), especially in form of NP which appears as a part of aspirin‐exacerbated respiratory disease (AERD). The aim of this study was to examine the concentrations of neuropeptides substance P (SP) and bradykinin (BK) in nasal secretions of patients with NP.MethodsFourteen patients with NP as a part of AERD with mild persistent asthma, 14 patients with NP without aspirin sensitivity, and 14 control subjects without nasal inflammation (C) entered this cross‐sectional study. Clinical parameters (symptoms, endoscopic, and radiological findings) were assessed. The concentrations of SP and BK were measured in the nasal secretion samples using commercial human enzyme immunoassay kits.ResultsThe concentration of SP in nasal secretions was significantly higher in NP patients without aspirin sensitivity and AERD patients compared to controls (p = .022; p < .0001, respectively), but higher in AERD than in non‐AERD patients (p = .018). The level of BK in nasal fluid was higher in non‐AERD and AERD NP patients than in controls (p < .0001; p < .0001, respectively), but also higher in AERD than in non‐AERD patients (p < .0001). We found high positive correlations between BK in nasal fluid and Lund–Mackay computed tomography (CT) score in both non‐AERD and AERD groups of NP patients.ConclusionOur results suggest more intense release of SP and BK from the nasal mucosa in patients with AERD than in patients with NP who do not have aspirin sensitivity. The strong correlation between concentration of BK in nasal secretions and CT score suggests that BK in nasal fluid could be used as a marker for disease severity as measured by the Lund–Mackay score.

Biodegradation of Organic Waste Using Bacillus Species Isolated From Soil

Organic waste can be enzymatically degraded by microbes. In this study, the Bacillus species were isolated from soil and identified as Bacillus subtilis, Bacillus licheniformis, Bacillus macquariensis, Bacillus brevis, and Bacillus circulans which were optimized considering pH (5, 7, 9) and temperature (37°C, 45°C, 55°C) for the maximum production of amylase, gelatinase, lipase, and cellulase, principally for the degradation of organic waste. The maximum production of amylase was found at 37°C with pH 7 and 9, gelatinase and lipase at 37°C with pH 5,7,9 by almost all identified species. Similarly, the production of cellulase was shown by Bacillus licheniformis only at 45°C, pH 5. The degradation was confirmed by the analysis of the solid content of degraded waste. The maximum degradation of starch and lipid-containing waste was shown by Bacillus macquariensis whereas Bacillus circulans were able to degrade gelatin-containing waste effectively. Bacillus species showed a synergistic effect in biodegradation. Bacillus subtilis and Bacillus licheniformis used in ratios 1:1 and 1:2 were found to be effective degraders of lipid and starch-containing waste respectively. Bacillus macquariensis, Bacillus brevis, and Bacillus circulans used in ratio 1:1:1 showed effective degradation of gelatin-containing waste. The degradation of the organic waste by multi-enzyme producer Bacillus species can be the most effective and eco-friendly method and their optimization for enzyme production can be beneficial for commercial enzyme production as well as for biotechnological applications. Int. J. Appl. Sci. Biotechnol. Vol 10(2): 104-111.

Fiber‐enriched biscuits prepared with enzyme‐treated corncob powder: Nutritional composition, physical properties, and sensory acceptability

Corncob (Zea mays L.) is a rich dietary fiber source, however, it is often neglected and has been mainly used as animal foodstuffs. In this study, corncob powder (CCP) was treated with Filtrase Sprint—a commercial enzyme preparation containing Talaromyces emersonii endo-1,3(4)-β-glucanase and endo-1,4-β-xylanase, to converse insoluble dietary fiber (IDF) into soluble dietary fiber (SDF), and then was added in a biscuit formulation. The suitable conditions for the enzymatic treatment were water content of 85% w/d.w., enzyme amount of 54 μl/g d.w., and 3 h incubation time. Biscuits cooperated with CCPs and increased significantly dietary fiber, total phenolic content, and antioxidant activity compared to the control sample. Enzyme-treated products resulted in lower hardness, higher fracturability, and higher overall acceptability than those incorporated with untreated counterparts. Novelty impact statement The results demonstrated a practical example of using enzymatic processing of corncob as a readily available high-fiber supplement for the human diet, which supports the concept of valorization of by-products from agriculture into high-value products and contributes to the sustainable development and the food security.

UV-B Protective and Antioxidant Activities of Protein Hydrolysate From Sea Cucumber (Holothuria scabra) Using Enzymatic Hydrolysis

Sea cucumber is rich in protein that can be used to prepare a potential derived bioactive peptide for antioxidant and protective effect against UV-B induced skin cell damage. This study aimed to optimize preparation of sea cucumber hydrolysate with both UV-B protective and antioxidant activities using three commercial enzymes using response surface methodology (RSM) with a face-centered central composite design (face- centered CCD). Hydrolysis time and concentration of enzyme effects on the degree of hydrolysis (DH), yield, antioxidant and UV-B protective activities of sea cucumber hydrolysates were determined. The optimum conditions for sea cucumber hydrolysis using papain (SCP), alcalase (SCA), or flavourzyme (SCF) were 3.6, 5.0, and 4.1% (w/w protein), respectively, and a hydrolysis time of 360 min. The resulting hydrolysates had a DH of 81-91%, yield of 13-14%, IC50 for DPPH radical scavenging activity of 0.3-4.1 mg/mL, FRAP of 0.5-0.6 mmol FeSO4/mL, and IC50 for ABTS radical scavenging activity of 1.3-1.6 mg/mL. The UV-B protective activity was reported as the HaCaT cell viability percentage after UV-B treatment. The SCP, SCA, and SCF hydrolysates showed 72.4, 74.5, and 71.3% cell viability, respectively. The concentration of hydrolysates with 80% survival of HaCaT cells was 0.21, 0.15 and 0.20 mg/mL for SCP, SCA and SCF, respectively. Thus, the SCP was selected for bioactive peptide isolation and characterization. The SCP contained hydrophilic and hydrophobic amino acids of 42.4 and 57.6%, respectively. The ultrafiltration and Sephadex G-25 gel filtration chromatography were done for peptide isolation from the SCP. Six potential peptides were identified using LC-MS/MS as Leu-Val-Asn-Glu-Leu-Thr-Glu-Phe-Ala-Gln (1163 Da), Leu-Val-Asn-Glu-Val-Thr-Glu-Phe-Ala-Gln (1149 Da), Phe-Val-Asp-Ser-Ser-Ala-Thr-Thr (826 Da), Phe-Asn-Asp-Leu-Gly-Ala-Trp (821 Da), Phe-Pro-Asp-Thr-Thr-Thr-Leu (793 Da), and Lys-Phe-Gly-Glu-Gly-Lys (664).

Production of protease with Bacillus megaterium DSM32: Partial characterisation of the enzyme and modelling of the production

Abstract Proteases hold an important position in today's world commercial enzyme market. Among various microbial producer genera, Bacillus is leading the commercial protease production. However, industry is still actively looking for new microbial protease producers with distinctive properties. Therefore, this study was undertaken for the evaluation of protease production by Bacillus megaterium DSM 32 strain in terms of its protease productivity, calculation of various production kinetics, partial characterisation of the enzyme, and modelling the protease production process. As results, the highest protease activity, specific cellular protease production rate, and protease productivity were calculated as 255.42 U mL−1, 36.2514 U g−1, and 16.1313 U mL−1 h−1, respectively, in shake flask fermentations. Partial characterisation studies showed that the enzyme has 45 °C and pH 8 as optimum working conditions, and its activity increased by 24% with the addition of 5 mM Mn+2 to the reaction medium. Additionally, the enzyme showed high stability and kept almost full activity in a cell-free medium for 20 days at 4 °C. Furthermore, modified Gompertz model provided the best fit in describing protease production with the lowest error and high fit values.

EVALUATION OF PHENOLIC COMPOUNDS IN WHITE WINES TREATED WITH ENZYMES

Wine's quality is influenced both by the grape characteristics and winemaking protocols. Awareness of the significant role of enzymes preparations in beverages technologies contributes to the optimization of the manufacturing process, for improving the chemical composition of the resulting wine and its organoleptic properties. This paper focuses on monitoring the impact of different commercial enzymes (pectinases and β-glycosides) on the main phenolic compounds content of Sauvignon blanc wines. For this experiment, ten phenolic compounds were quantified using a liquid-chromatography (LC) system coupled with ion trap mass spectrometer. The results indicated a significant influence of enzymes on wine’s phenolic fraction. Experimental samples presented high content in protocatechuic acid (9.99 - 13.75 μg/mL) and caftaric acid (2.69 - 9.80 μg/mL). The use of pectinases lead to an increase of phenolic compound’s concentration compared to the control.

Analysis and validation of silica-immobilised BST polymerase in loop-mediated isothermal amplification (LAMP) for malaria diagnosis

Bacillus stearothermophilus large fragment (BSTLF) DNA polymerase is reported, isolated on silica via a fused R5 silica-affinity peptide and used in nucleic acid diagnostics. mCherry (mCh), included in the fusion construct, was shown as an efficient fluorescent label to follow the workflow from gene to diagnostic. The R5 immobilisation on silica from cell lysate was consistent with cooperative R5-specific binding of R52-mCh-FL-BSTLF or R52-mCh-H10-BSTLF fusion proteins followed by non-specific protein binding (including E. coli native proteins). Higher R5-binding could be achieved in the presence of phosphate, but phosphate residue reduced loop-mediated isothermal amplification (LAMP) performance, possibly blocking sites on the BSTLF for binding of β- and γ-phosphates of the dNTPs. Quantitative assessment showed that cations (Mg2+ and Mn2+) that complex the PPi product optimised enzyme activity. In malaria testing, the limit of detection depended on Plasmodium species and primer set. For example, 1000 copies of P. knowlesi 18S rRNA could be detected with the P.KNO-LAU primer set with Si-R52-mCh-FL-BSTLF , but 10 copies of P. ovale 18S rRNA could be detected with the P.OVA-HAN primer set using the same enzyme. The Si-immobilised BSTLF outperformed the commercial enzyme for four of the nine Plasmodium LAMP primer sets tested. Si-R52-mCh-FL-BSTLF production was transferred from Cambridge to Accra and set up de novo for a trial with clinical samples. Different detection limits were found, targeting the mitochondrial DNA or the 18S rRNA gene for P. falciparum. The results are discussed in comparison with qPCR and sampling protocol and show that the Si-BSTLF polymerase can be optimised to meet the WHO recommended guidelines.Graphical abstract

Prevalence of Hepatitis B Infection Among Pregnant Women in Oman

ObjectiveThe primary objective of our study was to estimate the prevalence of hepatitis B infection among pregnant women in Oman. The secondary objectives were to test for other hepatitis B virus (HBV) serological markers and to determine hepatitis B viral load.MethodsIn this cross-sectional study conducted from June 2019 to December 2020, we randomly selected 2210 Omani women who attended antenatal clinics across the country. Pregnant women were tested for HBV surface antigen (HBsAg) using a commercial enzyme immunoassay; those who tested positive were further analyzed for other HBV serological markers: HBV core total antibody (anti-HBc), HBV core antibody IgM (immunoglobulin M) (anti-HBc IgM), hepatitis B virus e antigen (HBeAg) and hepatitis B virus e antibody (anti-HBe). They were also tested for hepatitis B viral load.ResultsThis study included 2210 women with a mean age of 39 years. Ninety-five percent of the women who were invited to participate consented and were included in the study. Thirty-three (1.49%) tested positive for HBsAg. All participants tested negative for HBeAg and anti-HBc IgM and positive for anti-HBc and anti-HBe, and 30 (90.9%) women had their hepatitis B viral load measured. Three (10.0%) had undetectable viral loads, 23 (76.7%) had low viral loads of < 2000 (IU/ml), 2 had moderate viral loads between 2000 and 200,000 (IU/ml) and one had a high viral load of 486,000 (IU/ml).ConclusionOur study shows that the HBV prevalence in pregnant women is 1.49%, which is lower than what was reported earlier. Nevertheless, antenatal screening is still warranted, as there are vital interventions for the newborn and the mother.

Thermophilic Geobacillus WSUCF1 Secretome for Saccharification of Ammonia Fiber Expansion and Extractive Ammonia Pretreated Corn Stover.

A thermophilic Geobacillus bacterial strain, WSUCF1 contains different carbohydrate-active enzymes (CAZymes) capable of hydrolyzing hemicellulose in lignocellulosic biomass. We used proteomic, genomic, and bioinformatic tools, and genomic data to analyze the relative abundance of cellulolytic, hemicellulolytic, and lignin modifying enzymes present in the secretomes. Results showed that CAZyme profiles of secretomes varied based on the substrate type and complexity, composition, and pretreatment conditions. The enzyme activity of secretomes also changed depending on the substrate used. The secretomes were used in combination with commercial and purified enzymes to carry out saccharification of ammonia fiber expansion (AFEX)-pretreated corn stover and extractive ammonia (EA)-pretreated corn stover. When WSUCF1 bacterial secretome produced at different conditions was combined with a small percentage of commercial enzymes, we observed efficient saccharification of EA-CS, and the results were comparable to using a commercial enzyme cocktail (87% glucan and 70% xylan conversion). It also opens the possibility of producing CAZymes in a biorefinery using inexpensive substrates, such as AFEX-pretreated corn stover and Avicel, and eliminates expensive enzyme processing steps that are used in enzyme manufacturing. Implementing in-house enzyme production is expected to significantly reduce the cost of enzymes and biofuel processing cost.

Enzymatically modified starch for paper surface sizing: Enzymes with different action modes and sites

Enzymatically modified starch is frequently applied as a kind of superior surface sizing agent to improve paper quality, but the available options for various enzyme types, except for α-amylases, are limited. Herein, starch was modified using various commercial enzymes with different action modes and sites, namely, α-amylase (CSTWA), β-amylase (CSTWB), pullulanase (CSTWP), glucoamylase (CSTWG), and compound enzyme (α-/β-amylase, CSTWAB), for sizing paper. The performance of the paper and the properties of the sizing agents were comprehensively determined. Moreover, the molecules of the modified starch were characterized to explore the enzymolysis mechanism. Results showed that enzymatic hydrolysis should be performed ahead of gelatinization. Compared with CSTWA, CSTWB and CSTWP endowed the paper with higher strength owing to their apposite viscosity and molecular mass distribution. CSTWG and CSTWAB contained excess low-molecular-mass molecules, making them weakly effective as surface-sizing agents. Therefore, pullulanase and β-amylase have better application prospects as surface-sizing agents than α-amylase.

Recovery of amino acids and peptides from pig bone soup using thermal pre‐treatment and enzymatic hydrolysis

Pig shoulder bones, by-products from the meat industry, are rich in protein and could be used to produce high amino acid products for food processing. In this study, after formulating the bone soup by heating it in water, we optimized the hydrolyzing process of the bone soup using three commercial enzymes (Flavourzyme, Protamex, and Alcalase). Alcalase hydrolysis gained the highest soluble protein recovery efficiency of 70.45% at the optimum condition, including hydrolysis temperature of 56°C, hydrolysis time of 138 min, and E/S ratio of 2.3%. The Alcalase hydrolysate achieved the peptide average molecular weight of 532 Da. In terms of nutritional value, the amino acid content gradually increased from the samples hydrolyzed with Alcalase, Flavourzyme, and Protamex (15.96, 16.60, and 20.71 mg/ml, respectively). However, the hydrolysate using Alcalase was the most favorable sample in sensory evaluation. Novelty Impact Statement Optimized soluble protein recovery of amino acids and peptides via enzymatic hydrolysis is essential to attain the highest possible nutritional value from raw materials like pig bone. Besides, thermal pretreatment and fat removal were implemented to identify soluble protein recovery efficiency (SPRE) precisely.

Enzyme-assisted extraction of anthocyanins and other phenolic compounds from blackcurrant (Ribes nigrum L.) press cake: From processing to bioactivities

The effects of commercial enzymes (pectinases, cellulases, beta-1-3-glucanases, and pectin lyases) on the recovery of anthocyanins and polyphenols from blackcurrant press cake were studied considering two solid:solvent ratios (1:10 and 1:4 w/v). β-glucanase enabled the recovery of the highest total phenolic content – 1142 mg/100 g, and the extraction of anthocyanins was similar using all enzymes (∼400 mg/100 g). The use of cellulases and pectinases enhanced the extraction of antioxidants (DPPH − 1080 mg/100 g; CUPRAC – 3697 mg/100 g). The freeze-dried extracts presented antioxidant potential (CUPRAC, DPPH), which was associated with their biological effects in different systems: antiviral activity against both non-enveloped viruses (enterovirus coxsackievirus A-9) and enveloped coronaviruses (HCoV-OC43), and cytotoxicity towards cancer cells (A549 and HCT8). No cytotoxic effects on normal human lung fibroblast (IMR90) were observed, and no anti-inflammatory activity was detected in lipopolysaccharides-treated murine immortalised microglial cells.

Efficiency of the removal of tetraethyl pyrophosphate (TEPP) pesticide in water: use of cork granules as a natural adsorbent on acetylcholinesterase activity in neuronal PC12 cell

Tetraethyl pyrophosphate (TEPP) is an organophosphate pesticide that irreversibly inhibits acetylcholinesterase (AChE). Cork powder or granules have been recommended as a sustainable sorbent to remove pesticides from water. In the present study, we evaluated the effectiveness of removing TEPP from water using wine corks to obtain cork granules as natural adsorbent, analyzing the TEPP effects on AChE activity in commercial enzyme from Electrophorus electricus and secreted by neuronal PC12 cells. TEPP inhibited AChE activity in a concentration-dependent manner. For the first time, we showed that different concentrations of TEPP diluted in water after adsorption experiments using cork granules decreased TEPP's inhibitory effects on AChE activity in commercial enzyme and neuronal PC12 cell culture medium. Our results suggest that the optimum removal of TEPP from water by corks was 91.4 ± 4.0%. Overall, the findings support the hypothesis that cork granules can be used to remediate pesticide-contaminated environments, such as those contaminated by organophosphate pesticides, and demonstrate a new application of a biochemical assay on AChE activity using a commercial enzyme or secreted by neuronal PC12 cells in culture as a possible methodologic strategy for evaluating the success of TEPP removal from water.

Recovery of cellulose and lignin from Eucalyptus by-product and assessment of cellulose enzymatic hydrolysis

In this study, a Eucalyptus by-product was fractionated into cellulose and lignin by pretreatment with NaClO2 to obtain α-cellulose, and hydrothermal pretreatment followed by sulfite-NaOH to obtain cellulose and lignin. The recovered celluloses were chemically and structurally characterized and enzymatically hydrolyzed using commercial enzyme preparations. The α-cellulose and cellulose treated with sulfite-NaOH had cellulose contents of 70.1% and 61.1%, respectively. α-cellulose hydrolyzed with Celluclast showed potential for cello-oligosaccharides formation, resulting in a maximum conversion of 30%, while Cellic CTec 2 resulted in 100% conversion to glucose, whereas cellulose treated with sulfite-NaOH showed potential for conversion to glucose with Cellulcast (44%) and Cellic CTec 2 (78%). The liquor from the sulfite-NaOH process resulted in a 20.8% yield of sulfonated lignin recovery. The use of different pretreatment techniques for the Eucalyptus by-product can be alternatives for the generation of products with high added value and commercial potential, such as cello-oligosaccharides, glucose, and sulfonated lignin.

Microbial production of multienzyme preparation from mosambi peel using Trichoderma asperellum.

Fruit and vegetable wastes create unhygienic conditions and pose a environmental pollution. The utilization of such wastes as carbon sources for production of enzyme with microbial intervention could be an ecofriendly and profitable approach, apart from diminishing the waste load. The present investigation focused on the feasibility of using mosambi (Citrus limetta) peel as substrate for multienzyme production (pectinase, cellulase and amylase) through microbial intervention. Fifteen fungi were isolated from organic waste and screened in vitro their potential of biodegradation of mosambi peel through enzymes production. The best performing isolate was selected and identified as Trichoderma asperellum NG-125 (accession number-MW287256). Conditions viz. temperature, pH, incubation time and nutrient addition were optimized for efficient enzymes production. The maximum enzyme activity (U ml-1min-1) of pectinase (595.7 ± 2.47), cellulase (497.3 ± 2.06) and amylase (440.9 ± 1.44) were observed at pH 5.5, incubation temperature of 30°C after 10days of fermentation. Moreover, macro-nutrients such as ammonium sulfate (0.1%) and potassium-di-hydrogen-ortho-phosphate (0.01%) further also enhanced the production of enzymes. The SDS-PAGE analysis of purified pectinase, cellulase and amylase using showed molecular mass of 43, 66 and 33kDa, respectively. The enzyme retention activity (ERA) of aforesaid enzymes was also tested with four different natural fiber matrices viz., bagasse, rice husk, paddy straw and wheat straw. Among these, the maximum ERA was observed on bagasse matrix (pectinase-56.35%, cellulose-77.68% and amylase 59.54%). Enzymatic juice clarification yield obtained with test enzyme was 75.8%, as compared to 80.5% of commercial enzyme. The result indicates that T. asperellum may be exploited as multifaceted biocatalysis.

MO735: Increased Serum Adipocyte Fatty Acid Binding Protein Level Associated with Endothelial Dysfunction in Patients Undergoing Hemodialysis

Abstract BACKGROUND AND AIMS Adipocyte fatty acid binding protein (A-FABP) is independently associated with atherosclerosis. Endothelial dysfunction is one of the causes of cardiovascular morbidity and mortality in patients with hemodialysis (HD). This study evaluated the correlation between serum A-FABP level and endothelial function measured by vascular reactivity index (VRI) in patients with HD. METHOD Fasting blood samples were obtained from 90 HD patients. A commercial enzyme immunoassay kit was used to measure A-FABP levels. The endothelial function and VRI were measured using a digital thermal monitoring (DTM) test (VENDYS). In this study, VRI &amp;lt; 1.0 was used as the poor vascular reactivity, 1.0 ≤ VRI &amp;lt; 2.0 was used as the intermediate vascular reactivity, and VRI ≥ 2.0 was used as the good vascular reactivity. RESULTS A total of 14 HD patients (15.6%) were categorized as poor vascular reactivity (VRI &amp;lt; 1.0), 38 HD patients (42.2%) were categorized as intermediate vascular reactivity (1.0 ≤ VRI &amp;lt; 2.0) and 38 HD patients had good vascular reactivity. Decreased pre-HD body weight (P = 0.016), post-HD body weight (P = 0.013), body mass index (P = 0.034), serum creatinine level (P = 0.010) and higher serum A-FABP level (P = 0.001) were associated with poor vascular reactivity. After multivariable forward stepwise linear regression analysis noted that serum creatinine level (β = 0.234, adjusted R2 change = 0.043; P = 0.019) was positively associated with, and serum A-FABP level (β = −0.347, adjusted R2 change = 0.152; P = 0.001) was negatively associated with VRI values in patients undergoing HD. CONCLUSION After adjusting for significant confounders, the serum A-FABP level was negatively correlated, and the serum creatinine level was positively correlated with VRI values in patients undergoing HD.