Enzymatic Hydrolysis of Porang by Streptomyces Violascens BF 3.10 Mannanase for the Production of Mannooligosaccharides

Porang (Amorphophallus muelleri Blume) is an indigenous Indonesian plant containing high hemicellulose as a source of glucomannan. An alternative way to produce a good quality of mannooligosaccharides was through hydrolysis of glucomannan by endo-β mannnase from actynomicetes. Based on 16S rRNA analysis, BF 3.10 isolate, isolated from Bukit Duabelas National Park soil, Jambi was identified as Streptomyces violascens BF 3.10. Reducing sugar was analyzed by dinitrosalicylic acid methods. The highest reducing sugar was achieved at the 72 hours of incubation. Mannanase of isolate BF 3.10 had the highest activity at pH 6 and temperature of 70 °C with enzyme activity of 16.38 U/mL and was stable at 4 °C for 48 h. During 5-hour of hydrolysis with substrate concentration of 0.25%, 0.5%, and 1% porang glucomannan dissolved in 10 mL enzyme, mannooligosaccharides were produced with the degree of polymerization of 2-3. Visualization of the products by using thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) methods showed that mannooligosaccharides produced comprised of glucose, mannobiose, mannotriose, and mannotetraose. The degree of polymerization and the simple sugars produced indicated that mannanase produced by S. violascens actively catalyzed the hydrolysis of 1.4-β-D-mannoside linkage from β-1.4-mannan backbone, that eventually produced simple sugars of mannooligosaccharides

Characterization of Bacterial Mannanase for Hydrolyzing Palm Kernel Cake to Produce Manno-oligosaccharides Prebiotics

Palm kernel cake (PKC) is a promising source of prebiotics, since it contains high amount of β-mannan which can be further hydrolyzed to manno-oligasaccharides (MOS), a prebiotic. Therefore, this research was carried out to analyze the capability of a bacterial isolate (A2 isolates) previously isolated from soils sample from around IPB campus to hydrolyze PKC. Based on 16S-DNA analysis, isolate A2 was identified as Brevibacillus borstelensis. Mannanase of A2 isolate had an optimum condition at 90 oC and pH 7. Mannanase activity of crude extracts using Locust Bean Gum (LBG) and PKC as substrates were 0.37U/mL and 0.032U/mL, respectively. However, the most favorable production of oligosaccharides based on the degree of polymerization was obtained after 72-h of incubation with the ratio of substrate:enzyme, 1.2:1, on 1.5% PKC as substrate. The manno-oligosaccharides prebio-tic obtained was found to interfere the growth of both lactic acid bacteria (Lactobacillus casei) and pathogenic microflora (Escherichia coli). E. coli apparently could not use this prebiotic as the carbon sources, in contrast to L. casei. Substitution of carbon source in medium with prebiotics reduced the capability of L. casei to produce organic acids. It is concluded that local A2 isolate (B. borstelensis) produces mannanase which can be used to produce prebiotics from PKC

Characterization of Three Benzoate Degrading Anoxygenic Photosynthetic Bacteria Isolated From the Environment

Three anoxygenic photosynthetic bacteria, DS-1, DS-4 and Cas-13, have been examinated for themorphological and physiological properties. All strains were rod-shape cells with a swollen terminal endGram negative, motile, non-halophilic, non-alkalophilic and non-acidophilic, and capable of utilizinbenzoate aerobically and photo-anaerobically. Sequence analysis of part of 16S rRNA genes showed that DS1 and Cas-13 were closely related to Rhodopseudomonas palustris Strain 7 with a similarity of 97%, whereaDS-4 may not be closely related to the former two strains with a similarity of 78% based on the constructephylogenic tree. Spectral analysis indicated that the three bacteria had bacteriochlorophyl a and normaspirilloxanthin series. Growth in medium enriched with vitamin and supplemented with benzoate as their sole C-sources wabetter than in medium without vitamin. Benzoate degradation in medium with vitamin was accelerated. Thability to grow on benzoate without added vitamins indicated that the bacteria were able to synthesize theown vitamins

Effect of Dietary Mannan-oligosaccharides From Copra Meal on Intestinal Microbes and Blood Profile of Broiler Chickens

Mannan-oligosaccharides (MOS) is a common prebiotics used as a feed additives. The use of antibiotic can be replaced by MOS to reduce the number of pathogenic bacteria in small intestines of chickens. The objective of this study was to determine the effect of MOS extracted from copra meal on intestinal microbial population and blood profile of broiler chickens. The study was conducted by using 150 day-old male broiler chickens (average body weight 46.00 + 1.41 g). The design of this experiment was a completely randomized design (CRD) with 3 dietary treatments, 5 replications and 10 birds in each replication. The dietary treatments consisted of control diet (corn-soybean meal diet without supplementation of antibiotic and prebiotic), control + 0.1% antibiotic bambermycin, and control + 0.035% MOS. For the first 4 weeks the birds were fed treatment diets, followed by commercial feed for 2 weeks. The parameters observed in this study were intestinal microbes population and blood profiles. The results showed that the administration of 0,035% MOS increased the lactic acid bacteria (LAB) number compared with antibiotics feed (P0.05) with the control group. The total aerobic and coliform bacteria were higher in MOS and antibiotic treatment compared than control group. The supplementation of 0.035% MOS from copra meal in diet increased the level of leucocytes which is important to immune response of broilers and its USAge was safe because the level of chicken blood profile were within the normal range

Xylanase Activity of Streptomyces violascences BF 3.10 on Xylan Corncobs and its Xylooligosaccharide Production

Corn is one of the important carbohydrate sources in Indonesia that is mainly used for food and industrial materials. In addition, the byproducts of corn, such as corncobs, have been reported as xylan-containing materials that can be utilized as substrate in xylooligosaccharides (XOS) production. XOS are natural prebiotic fibers that can enhance the performance of animal’s digestive system. The main objective of this study was to exploit xylan from corncobs to produce XOS. The research consisted of extraction and production of xylan from corncobs and the synthesis of XOS from corncob-produced xylan. The corncob and Streptomyces violascens BF 3.10 xylanase is a collection of PPSHB IPB Laboratory. Corncobs xylan extracted by using alkaline method and reducting sugar was analyzed by dinitrosalicylic acid method. The xylan extraction from corncobs could produce 7.93% (w/w) of xylan. The activity of S. violascens BF 3.10 xylanase on the substrate of concorb-produced xylan was 6.4 U/mL at the optimum temperature of 60 °C in 50 mM phosphate buffer with pH 5.5. The thin layer chromatography analysis indicated that 1% (w/v) corn-cob xylan could produce XOS with degree of polymerization (DP) 3.92.  XOS, with DP ranging from 2-4, could be used as a livestock feed mixture to stimulate the growth of normal microbes in the gastrointestinal tract of livestock.Key words: corncobs, Streptomyces violascens BF 3.10, xylan, xylanas

Using Streptomyces Xylanase to Produce Xylooligosacharide From Corncob

Streptomyces 234P-16 and SKK1-8 are xylanase-producing bacteria. Corncob xylan were extracted using acidified method. Crude enzymes (produced by centrifuging the culture) were used to hydrolyze xylan from 2 varieties of corncob. Crude extract activity was measured by using DNS (Dinitrosalisilic Acid) method. Xylanase from strain 234P-16 has the highest activity if cultivated in 1% Hawaii xylan, whereas strain SKK1-8 on 1.5% Bisma xylan. SKK1-8 xylanase can hydrolize corncob xylan (1% Hawaii or 1.5% Bisma xylan) within 4 hours and produce xylooligosacharide with polymerization degree of 4.76 and 6.37, respectively

Purifikasi Dan Karakterisasi Enzim Pektinase Dari Aspergillus Ustus Bl5 [Purification and Characterization of Pectinase From Aspergillus Ustus Bl5]

Pectinase is an enzyme that could hydrolyze pectin into galacturonic acid. Natural pectinase was produced by microbes such as bacteria, yeast, fungi and Actinomycetes. Application of pectinase in industry were mainly in juice industry, textile, pulp, tea, cocoa and coffee fermentation. In this research, we conducted purification and characterization of pectinase produced by Aspergillus ustus BL5 in submerged fermentation using commercial pectin. The result showed that the optimum of pectinase production was reached at 120 hours fermentation process with specific activity 0.59 U/mg. The crude extract of pectinase was then concentrated using PEG 6000 and purified by Sephadex G-75 gel filtration chromatography. There were 2 fractions contained pectinase which the activity was 4.15 U/mg (pectinase A) and 3.3 U/mg (pectinase B), respectively. Compare to crude extract, the yield product of pectinase A and B increased 6.94 and 5.53 times, respectively. The purified pectinase A have optimum temperature at 50 oC and optimun pH at 5

Xylanase Activity of Streptomyces violascences BF 3.10 on Xylan Corncobs and its Xylooligosaccharide Production

Corn is one of the important carbohydrate sources in Indonesia that is mainly used for food and industrial materials. In addition, the byproducts of corn, such as corncobs, have been reported as xylan-containing materials that can be utilized as substrate in xylooligosaccharides (XOS) production. XOS are natural prebiotic fibers that can enhance the performance of animal’s digestive system. The main objective of this study was to exploit xylan from corncobs to produce XOS. The research consisted of extraction and production of xylan from corncobs and the synthesis of XOS from corncob-produced xylan. The corncob and Streptomyces violascens BF 3.10 xylanase is a collection of PPSHB IPB Laboratory. Corncobs xylan extracted by using alkaline method and reducting sugar was analyzed by dinitrosalicylic acid method. The xylan extraction from corncobs could produce 7.93% (w/w) of xylan. The activity of S. violascens BF 3.10 xylanase on the substrate of concorb-produced xylan was 6.4 U/mL at the optimum temperature of 60 °C in 50 mM phosphate buffer with pH 5.5. The thin layer chromatography analysis indicated that 1% (w/v) corn-cob xylan could produce XOS with degree of polymerization (DP) 3.92. XOS, with DP ranging from 2-4, could be used as a livestock feed mixture to stimulate the growth of normal microbes in the gastrointestinal tract of livestock

Damping-off disease reduction using actinomycetes that produce antifungal compounds with beneficial traits

Actinomycetes are considered to be the biggest producer of bioactive compounds which are expected to have antifungal activity for controlling many fungi such as Rhizoctonia solani. The objective of this study was to obtain potential soybean rhizosphere actinomyc- etes as a biocontrol agent for R. solani which cause damping-off disease both in vitro and in vivo, including their ability to produce siderophore, chitinase, and HCN. Out of 26 isolates, 18 (56%) showed diverse antifungal activities against R. solani with percentages of inhibi- tion radial growth (PIRG) from 18.9 to 64.8%, as evaluated by a dual culture method. Ten isolates with the strongest antifungal activity were numbered for further characterization. All the tested isolates were not antagonistic towards Bradyrhizobium japonicum. These iso- lates were able to suppress damping-off disease caused by R. solani in the greenhouse exper- iment. Isolate ASR53 showed the highest disease suppression, 68% and 91% in sterile and non-sterile soil, respectively. Based on 16S rRNA sequence analysis this isolate belonged to Streptomyces violaceorubidus LMG 20319 (similarity 98.8%) according to GenBank data base available at www.ncbi.nlm.gov.nih. Furthermore, isolate ASR53 had significantly longer roots and shoots, as well as greater fresh and dry weights of seedlings than the con- trol. Crude extract derived from ASR53 isolates contained 10 dominant compounds that were biologically active against fungal pathogens. Thus, this study suggests that the applica- tion of potential actinomycetes of the soybean rhizosphere can act as a promising biocon- trol agent against damping-off disease caused by R. solani