Hidrolisis Protein Teripang Hitam (Holothuria edulis) Menggunakan Bromelin Kasar dari Batang Nanas (Ananas comocus L)

Black sea cucumber is one type of sea cucumber whose protein content is quite high at 74-82%, not yet widely used by the community as a source of protein. One way to use it is to process the protein into a protein hydrolyzate using the bromelain enzyme which was isolated from pineapple stems. This study aims to determine the concentration of the bromelain enzyme extract and the optimum hydrolysis time to the value of the hydrolysis degree of the hydrolyzate of black sea cucumber protein. Protein hydrolysates are produced by variations of the crude bromelain concentration of 9%, 10%, 11%, and 12% (v/v) and hydrolysis time of 120, 180, 240, and 300 minutes. The results showed that the optimum degree of hydrolysis was obtained with the use of bromelain extract with a concentration of 12% with a value of 11.43% and a hydrolysis time of 240 minutes with a value of 19.17%.

Extracting Sweetening and Bioactive Compounds From Stevia Rebaudiana Using Cellulase Enzyme

This study was conducted to find out the sweetener and bioactive compounds in the Stevia plant’s enzyme extract and its antioxidant and antibacterial effect. The enzyme extract was used in a ratio of 1:15 (w: v) with the use of sodium phosphate buffer solution (pH=4) in an equal mixing ratio with the enzyme. Extraction was carried out at 55°C with a time of 25 minutes to extract and determine steviol glycosides using HPLC. Stevioside and Rebaudioside-A (5.101 and 3.027 mg/g) were obtained, respectively. The study showed that Stevia contains high amounts of phenols and flavonoids (83.052 and 71,765) mg/ml, respectively. The enzyme extract gave an antioxidant activity of 71.367% compared to BHT which gave an activity of 77.267%. It gave the highest inhibitory activity against Bacillus subtilis, which was 14 mm, followed by Staphylococcus aureus and E. coli. The bioactive compounds were diagnosed by GC-MS and contained important bioactive compounds such as Hydroxydehydrostevic acid (steviol), 1,2-Benzenediol, 4-Ethylcatechol, 9-Octadecenamide, (Z)-, Isosteviol methyl ester, gamma-Sitosterol.

Kinetic Analysis of Crude Enzyme Extract Produced Via Solid State Fermentation of Banana Pseudo Stem Waste

Banana Pseudo stem waste after each harvest contributes about 70–80 Milli Tons Per hector. The banana pseudo stem will be thrown as waste biomass after each harvest as it is unstable for the upcoming harvest. The biggest challenge in banana cultivation is the utilization of biomass of banana pseusostem waste into valuable products. In this study, Xylano-pectinase enzyme extract was produced from banana pseudo stem waste under solid-state fermentation by Enterobacter cloacae PMC04. The highest pectinase and xylanase activities obtained using banana pseudo stem as carbon source were 124.62 U/ml and 173.81 U/ml respectively. Thermodynamics stated that range 40-50oC were considered to be the optimal temperature for xylano-pectinase enzyme production and subsequent degumming of banana fibers. The crude enzyme extract were then used in the degumming of banana fibers for textile application. Textile processing of banana fiber necessitates the removal of hemicellulose substance which can be achieved by crude xylano-pectinase enzyme. It was found that crude xylano-pectinase was efficient in the removal of hemicellulose substance from the fibers. Results obtained from this study demonstrate that the proposed bioprocess could be successfully applied for the degumming of banana fibers sustainably.

Enzymatic Hydrolysis of Lignocellulosic Biomass Using an Optimized Enzymatic Cocktail Prepared From Secretomes of Filamentous Fungi Isolated From Amazonian Biodiversity

The use of lignocellulosic biomass (LCB) has emerged as one of the main strategies for generating renewable biofuels. For the efficient use of such feedstock, pretreatments are essential. The hydrolysis of cellulose – major component of LCB - demands enzymatic cocktails with improved efficiency to generate fermentable sugars. In this scenario, lignocellulolytic fungi have enormous potential for the development of efficient enzyme platforms. In this study, two enzymatic cocktails were developed for hydrolysis of two lignocellulosic biomasses: industrial cellulose pulp and cassava peel. The solid biomass ratio in relation to the protein content of the enzyme cocktail were performed by experimental design. The optimized cocktail for the hydrolysis of cellulose pulp (AMZ 1) was composed, in protein base, by 43% of Aspergillus sp LMI03 enzyme extract and 57% of T. reesei QM9414, while the optimal enzyme cocktail for cassava peel hydrolysis (AMZ 2) was composed by 50% of Aspergillus sp LMI03 enzyme extract, 25% of the extract of P. citrinum LMI01 and 25% of T. reesei. The ratio between solids and protein loading for AMZ 1 cocktail performance was 52 g/L solids and 30mg protein/g solids, resulting in a hydrolytic efficiency of 93%. For the AMZ 2 cocktail, the hydrolytic efficiency was 78% for an optimized ratio of 78g/L solids and 19mg protein/g solids. These results indicate that cocktails formulated with enzymatic extracts of P. citrinum LMI01, Aspergillus sp LMI03 and T. reesei QM9414 are excellent alternatives for efficient hydrolysis of plant biomass and for other processes that depend on biocatalysis.

Valorization of Lignocellulosic Wastes to Produce Phytase and Cellulolytic Enzymes from a -Thermophilic Fungus, Thermoascus aurantiacus SL16W, under Semi-Solid State Fermentation

Agricultural wastes are lignocellulosic biomasses that contain high mineral and nutrient contents. This waste can be used as a raw material in industrial enzyme production by microbial fermentation. Phytase is an important enzyme used in animal feed to enhance the amount of phosphorus available for the growth and overall health improvement of monogastric animals. Fungi offer high potential as an effective source in the production of various extracellular enzymes. In this study, the production of lignocellulolytic enzymes (endoglucanase and xylanase) and phytase by a thermophilic fungus, namely Thermoascus aurantiacus strain SL16W, was evaluated using sixteen different Thai agricultural forms of waste under conditions of high temperature (45 °C). Semi-solid state fermentation was used in the production experiments. The results of this study reveal that the highest phytase activity (58.6 U/g substrate) was found in rice bran, whereas the highest degrees of activity of endoglucanase and xylanase were observed in wheat bran and red tea leaves at 19 and 162 U/g substrate, respectively. Consequently, the optimal conditions for phytase production of this fungus using rice bran were investigated. The results indicate that the highest phytase yield (58.6 to 84.1 U/g substrate) was observed in rice bran containing 0.5% ammonium sulfate as a nitrogen source with 10 discs of inoculum size at a cultivation period of 9 days at 45 °C and moisture content of 95%. Notably, the phytase yield increased by 1.71-fold, while endoglucanase and xylanase were also increased by 1.69- and 1.12-fold, respectively. Furthermore, the crude enzyme obtained from the optimal condition was extracted. The crude enzyme extract was then separately added to red tea leaves, rice straw, corncobs, palm residue, and peanut husks. Subsequently, total reducing sugar and phosphorus contents were determined. The results indicate that the highest level of reducing sugar (122.6 mg/L) and phosphorus content (452.6 mg/L) (p < 0.05) were obtained in palm residue at 36 and 48 h, respectively, after the addition of the crude enzyme extract. This study has provided valuable information on a potentially eco-friendly way to valorize agricultural waste into value-added products as industrial enzymes.

Response of albino rats fed varying levels of enzyme supplemented tigernut meal based diets.

A 21-day feeding trial to assess the effect of replacing 0,10,20,30 and 40% maize with enzyme supplemented tigernut meal on the performance, nutrient digestibility, haematological traits, serum chemistry and carcass yield of one hundred and eight albino rats of the wistar strain was conducted in a complete randomised design. Nine experimental diets (1,2,3,4,5,6,7,8 and 9) were formulated in all. Diet 1 contained 40% maize + 0% tigernut meal without enzyme supplementation. Diets 2,3,4.5 and 6,7,8,9 were constituted by replacing maize at 10.20,30 and 40% levels with tigernut meal supplemented with 0.05% roxzyineG® and enzyme extract respectively. The results revealed that of all the parameters considered, only the gut characteristics were not significantly (P>0.05) influenced by the dietary treatments. Diets 5 and 9 (40% enzyme supplemented tigernut ineal) proved to be the best by having the best performance and nutrient digestibility coefficients. It was then concluded that 40% inclusion level of enzyme supplemented tigernut meal as a replacement for maize proved to be better in rats than dicts that contained 0, 10, 20 and 30% levels of enzyme supplemented tigernul meal based diets.

Optimization of enzymatic hydrolysis for bioethanol production by semi-simultaneous saccharification and fermentation using mature coconut fibre

Alternative substrates to produce useful chemicals such as biofuel have been attractive, in particular, for cellulosic ethanol production. In this context, the objective of this work was optimized the synergistic mixture of enzymes and bioethanol production. The enzymes of Trichoderma reesei and crude enzyme extract from Lichtheimia ramosa were used in the hydrolysis of mature coconut fibre pretreated by sequential process of alkaline hydrogen peroxide (Alk-H2O2)-sodium hydroxide (NaOH). Furthermore, these enzymes and pretreated vegetable biomass were applied in the bioethanol production by Saccharomyces cerevisiae in semi-simultaneous saccharification and fermentation strategy (SSSF). Resulting in the yields and conversions of delignified mature coconut fibre into reducing sugars between 12.7-82.14% and 0.09-0.64 g reducing sugars/g dry biomass, respectively, with an initial hydrolysis rate at 12 h between 0.10-0.89 g/(L.h). Yields and conversions of delignified mature coconut fibre into glucose between 10.16-83.78% and 0.06-0.43 g glucose/g dry biomass, in that order, with an initial hydrolysis rate at 12 h between 0.03-0.35 g/(L.h). Bioethanol production by S. cerevisiae using delignified mature coconut fibre, enzymes from T. reesei and crude enzyme extract from L. ramosa resulted in the production of 4.62 g/L, yield of 0.41 g ethanol/g glucose and volumetric productivity of ethanol of 0.13 g/(L.h), respectively. The results showed synergistic effects between enzymes from T. reesei and crude enzyme extract from L. ramosa, without promoting inhibition in the alcoholic fermentation. Therefore, allowing to formulate an optimized enzymatic preparation aiming cellulosic ethanol production.

Application of Biological Pretreatment using Enzyme Extracts from Filamentous Fungi to Improve Ruminal Degradability of Oil Palm Fronds.

Background Malaysia is one of the largest producer of palm oil in the world with yearly yield of 19 000 million tonnes. With its large scale production of palm oil, Malaysia produces approximately 30 million tonnes of oil palm fronds (OPF) annually. The OPF as the main agricultural by-product in Malaysia have limited use especially as ruminant feed. It is due to its poor degradability which is caused by its high lignin content. One of the most promising method to increase the utilization of OPF as ruminant feed is via biological pretreatment. Previous study has shown an increase of 12% of ruminal degradability when pretreated using enzyme extract from white rot fungi Results The isolation of fungi yielded 11 fungi isolates. Upon the determination of their laccase, manganese peroxidase, lignin peroxidase, carboxylmethylcellulase, avicelase and xylanase, fungi F1, F2 and F4 were selected as the fungi with desirable enzyme activity. The fungi F1, F2 and F4 were then identified as Trichoderma harzianum MK027304, Trichoderma harzianum MK027305 and Fusarium solani MK027307 respectively. The highest total gas production was produced by OPF pretreated with enzyme extract from 15 days of solid state fermentation of Trichoderma harzianum MK027304. The highest methane was produced by OPF pretreated with enzyme extract from 30 days of solid state fermentation of Trichoderma harzianum MK027305. Highest volatile acid produced by OPF pretreated with enzyme extract from Fusarium solani MK027307 after 45 days of solid state fermentation. The apparent rumen degradable carbohydrate of the pretreatment also showed the highest increase over the unpretreated OPF with increment of 35.29%. Conclusions The pretreatment using enzyme extract from 45 days of solid state fermentation of Fusarium solani MK027307 have shown the ability to increase the degradability of OPF by 35.29%. This finding may decrease the feeding cost of ruminant by increasing the effectiveness of utilization of OPF.