scholarly journals PRODUKSI ENZIM KITINASE DARI Aspergillus niger MENGGUNAKAN LIMBAH CANGKANG RAJUNGAN SEBAGAI INDUSER

2016 ◽  
Vol 1 (1) ◽  
pp. 34 ◽  
Author(s):  
Purkan Purkan ◽  
Afaf Baktir ◽  
Arju Rohmah Sayyidah
Keyword(s):  
Enzyme Activity ◽  
Aspergillus Niger ◽  
Cell Mass ◽  
Hydrolytic Enzyme ◽  
Ph Optimum ◽  
Fermentation Time ◽  
Activity Test ◽  
Crab Shell Waste ◽  
Dry Cell ◽  
Local Nature

AbstrakKitinase merupakan enzim hidrolitik yang dapat menghidrolisis kitin pada ikatan β-1,4-glikosidiknya dengan menghasilkan derivat-derivat kitin seperti oligomer kitin yang mempunyai banyak manfaat. Penelitian ini bertujuan untuk melakukan pengembangan produksi enzim kitinase dari sumber lokal yang melimpah di alamserta murah dengan melakukan optimasi substrat dalam hal ini digunakan substrat tetes tebu (molase) dan limbah cangkang rajungan untuk produksi enzim kitinase dari Aspergillus niger. Sebelumnya, dilakukan kultivasi isolat kapang Aspergillus niger dengan membuat kurva pertumbuhan menggunakan metode masa sel kering dimana dari hasil penelitian inokulasi optimal adalah 22 jam. Pada proses produksi, diperoleh waktu fermentasi optimal adalah 52 jam dengan menentukan uji aktivitasnya menggunakan metode turbidimetri. Hasil optimasi substrat menunjukkan bahwa enzim kitinase yang maksimal diperoleh pada penambahan molase 0,5% (b/v) dengan unit aktivitas enzim 0,14726 (U/mL) dan cangkang rajungan 2% (b/v) dengan unit aktivitas enzim yang dihasilkan 0,12826 (U/mL). Kitinase dari Aspergillus niger ini mempunyai pH optimal 6 dan suhu optimal 40 oC. Kata kunci: Aspergillus niger, kitinase, cangkang rajungan, molase   AbstractChitinase is a hydrolytic enzyme that hydrolyzes chitin on β-1,4-glycosidic bond and thereby producing chitin derivatives such as chitin oligomers that have multiple benefits. The purpose of this research was to develop the production of chitinase enzyme from cheap and are abundant local nature sources, by optimizations substrate in this case the substrate used molasses and crab shell waste for the production of chitinase enzyme from Aspergillus niger. Previously, isolates of Aspergillus niger cultivated by creating a growth curve using dry cell mass method which from the results of research inoculation optimal are 22 hours. In the production process, obtained the optimum fermentation time is 52 hours to determine the activity test using turbidimetry method. Result of substrate optimizations indicate that chitinase enzyme maximum by addition of molasses obtained in 0.5% (w/v) with enzyme activity units 0.14726 (U/mL) and crab shells 2% (w/v) with enzyme activity units 0.12826 (U/mL). Chitinase from Aspergillus niger has a pH optimum 6 and temperature optimum 40 oC. Keywords: Aspergillus niger, chitinase, crab shells, molasses

scholarly journals Produksi Enzim Selulase olehAspergillus niger pada Ampas Sagu

2015 ◽  
Vol 16 (1) ◽  
pp. 1 ◽  
Author(s):  
Nora Idiawati ◽  
Elliska Murni Harfinda ◽  
Lucy Arianie
Keyword(s):  
Enzyme Activity ◽  
Aspergillus Niger ◽  
Moisture Content ◽  
Cellulase Enzymes ◽  
Fermentation Time ◽  
Cellulase Enzyme ◽  
Sago Waste ◽  
Solid Fermentation

Production of cellulase by Aspergillus niger was carried out by growing the cultureson sago waste. Sago waste containscellulose that has not been used optimally. Cellulose is a polysaccharide consisting of glucose monomers linked by β-1,4-glycosides bonds. Glycoside bonds in cellulose can be enzymatically hydrolyzed into glucose with cellulase enzymes. Solid fermentation used to produce cellulase on sago waste as substrate was influenced by pH (3 to 6), moisture content(40% to 85%), and fermentation time (4 to 10 days). Products of the cellulase enzyme activity was measured by phenolsulfuricacid method. The results showed that the highest cellulase enzyme activity was 0.172 U/mL obtained at 85%moisture content, pH 5, and 8 days of fermentation time.

scholarly journals Cellulase Enzyme Production From Rice Straw Using Solid State Fermentation and Fungi Aspergillus niger ITBCC L74

2018 ◽  
Vol 156 ◽  
pp. 01010 ◽  
Author(s):  
Siti Maftukhah ◽  
Abdullah Abdullah
Keyword(s):  
Enzyme Activity ◽  
Aspergillus Niger ◽  
Protein Content ◽  
Rice Straw ◽  
Lignin Content ◽  
Enzyme Characterization ◽  
Raw Material ◽  
Enzymatic Reactions ◽  
Fermentation Time ◽  
Cellulase Enzyme

Rice straw is one of very abundant waste of agricultural and has not utilized maximally. This waste contain cellulose and potential in the manufacture of cellulase enzymes. Research on the production of cellulase enzyme from lignocellulose has been done a lot of enzyme activity is still low. This research using cellulose is 71.95% and conducted with 6 stages. First, the preparation of raw material. Second, the decrease of lignin content with alkali pretreatment. Third, the breeding of fungi Aspergillus niger ITBCC L74 . Fourth, incubation in the inoculum. Fifth, the production of cellulase enzyme by solid fermentation method. Finally, the analysis includes protein content, enzyme activity, enzyme characterization and kinetics of enzymatic reactions. The highest enzyme activity of this study is 3.12 U/ml and protein content is 0.34 mg/ml with fermentation time is 4 day and water content is 75%. In enzyme characterization obtained optimum pH and temperature are 4 and 60°C, respectively. And obtained paramatic kinetic are Vmax and Km for 40, 50, 60 and 70°C temperature are Vmax: 6.42; 4.7; 5.82 and 4.46 U/ml and Km : 1.32; 0.38; 0.32; and 0.12%, respectively.

scholarly journals OPTIMASI PROSES UNTUK EKSPRESI GEN ENDOGLUKANASE DARI Bacillus sp. RP1 OLEH Escherichia coli BL21 (DE3)/ egc

2018 ◽  
Vol 5 (1) ◽  
pp. 44
Author(s):  
Hans Victor ◽  
Maelita Ramdani Moeis
Keyword(s):  
Escherichia Coli ◽  
Enzyme Activity ◽  
Process Optimization ◽  
Rice Hull ◽  
Bacillus Sp ◽  
Fermentation Time ◽  
E Coli ◽  
Cell Weight ◽  
Dry Cell Weight ◽  
Dry Cell

Process Optimization for Endoglucanase Gene Expression Derived from Bacillus sp. RP1 by Escherichia coli BL21 (DE3)/egcABSTRACTCellulases are one of the most used enzymes in industrial processes. In an effort to increase production, industries have developed strategies such as isolating new cellulase producing strains, genetic engineering and process optimization since the last 50 years. One endoglucanase producing strain, Bacillus sp. RP1 was isolated from hot springs. The ribosome binding site and coding sequence of the endoglucanase gene (egc) from Bacillus sp. RP1 was cloned into pGEM-T Easy. The recombinant plasmid was used to transform E. coli BL21 (DE3). Cloning was followed by process optimization. Medium composition was selected using Plackett-Burman design. The medium components tested were rice hull, molasses, ammonium chloride, urea and fishmeal. Rice hull and molasses were found to be the factors most influencing enzyme activity and dry cell weight, respectively. The next step involved Box-Behnken method and response surface methodology to optimize the responses against molasses concentration, rice hull concentration and fermentation time. The concentration intervals used to test were 1%, 5.5% and 10% while the fermentation time used were 24, 36 and 48 hours. The conditions which optimized both enzyme activity and dry cell weight were 7.45% molasses, 6.45% rice hull and 39.52 hours of fermentation.Keywords: Bacillus sp. RP1, E. coli BL21 (DE3), egc, Endoglucanase, optimization ABSTRAKSelulase adalah salah satu enzim yang banyak dimanfaatkan dalam berbagai industri. Sebagai upaya untuk memenuhi kebutuhan, 50 tahun terakhir dikembangkan beberapa strategi untuk meningkatkan produksi selulase yang mencakup rekayasa genetika dan optimasi proses. Karena itu, dilakukan kloning gen egc dan RBS yang berasal dari Bacillus sp. RP1 yang diisolasi dari sumber air panas ke dalam vektor pGEM-T Easy. E. coli BL21 (DE3) ditransformasikan dengan vektor yang mengandung gen egc tersebut. Setelah kloning, optimasi proses berupa desain medium turut dilakukan untuk mengoptimalkan ekspresi gen egc. Desain medium diawali dengan seleksi komposisi medium menggunakan metode Plackett-Burman. Komponen medium yang diuji adalah kulit beras, molase, amonium klorida, urea dan tepung ikan. Kulit beras dan molase diperoleh sebagai bahan yang paling berpengaruh terhadap aktivitas enzim dan berat kering sel. Tahap selanjutnya melibatkan metode statistik Box-Behnken dan metodologi respons permukaan yang bertujuan mengoptimalkan respons aktivitas enzim dan berat kering sel terhadap konsentrasi molase, konsentrasi kulit beras dan lama fermentasi. Konsentrasi yang diuji adalah 1%, 5,5% dan 10%, sedangkan lama fermentasi yang diuji adalah 24, 36 dan 48 jam. Konsentrasi optimal molase adalah 7,45% dan konsentrasi optimal kulit beras adalah 6,45% dengan lama fermentasi optimal 39,52 jam.Kata Kunci: Bacillus sp. RP1, E. coli BL21 (DE3), egc, Endoglukanase, optimasi

Optimization of medium and cultivation conditions forl-amino acid oxidase production byAspergillus fumigatus

2009 ◽  
Vol 55 (9) ◽  
pp. 1096-1102 ◽  
Author(s):  
Susmita Singh ◽  
B. K. Gogoi ◽  
R. L. Bezbaruah
Keyword(s):  
Enzyme Activity ◽  
Amino Acid ◽  
Cell Mass ◽  
Industrial Applications ◽  
High Yield ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
North East ◽  
The North ◽  
Dry Cell

A fungal strain was selected from the microbial repository of the North-East Institute of Science and Technology, Jorhat, India, which could produce a high yield of l-amino acid oxidase. 18SrRNA, ITS1, 5.8SrRNA ITS2, and partial 28 S rRNA sequencing and phenotypic characteristics indicate that it belong to the species Aspergillus fumigatus (designated as P13). Maximum production of enzyme (59.55 × 10−3 U/mg dry cell mass) was obtained in a medium containing 10 g/L glucose, 4 g/L yeast extract, and 4 g/L ammonium sulfate, with 20 mmol/L of l-threonine as the inducer. The optimum temperature for enzyme production was 30 °C at pH 7.0, with a shaking speed of 200 r/min. At 96 h, the enzyme activity was maximum. The A. fumigatus P13 l-amino acid oxidase accepts a broad substrate range, and the maximum enzyme activity (20.41 × 10−3 U/mg dry cell mass) was obtained with 50 mmol/L of dl-tyrosine. In the literature, no reports have been found regarding the production of l-amino acid oxidase by A. fumigatus. The enzyme showed enantiomerically pure amino acid formation, which has tremendous demand in industrial applications.

Pyruvate carboxylase of Aspergillus niger: kinetic study of a biotin-containg carboxylase

1969 ◽  
Vol 47 (7) ◽  
pp. 697-710 ◽  
Author(s):  
Helen A. Feir ◽  
Isamu Suzuki
Keyword(s):  
Enzyme Activity ◽  
Aspergillus Niger ◽  
Pyruvate Carboxylase ◽  
Active Sites ◽  
Kinetic Studies ◽  
Kinetic Mechanism ◽  
Product Inhibition ◽  
Ph Optimum ◽  
Metal Cofactor ◽  
Michaelis Constants

Pyruvate carboxylase was partially purified from Aspergillus niger and the properties were studied. The enzyme was found to be cold-labile and protected by 25% glycerol. The pH optimum was determined to be 7.9–8.0. The enzyme was shown to be a biotin-containing enzyme by its inactivation by avidin and protection against such inactivation by biotin. The enzyme activity was stimulated by K+ ions and inhibited by Na+ ions. Acetyl-CoA had no effect on enzyme activity, but L-aspartate was inhibitory. Apparent Michaelis constants were determined for the substrates and metal cofactor involved, i.e. pyruvate, ATP, bicarbonate, and Mg2+.Initial-velocity studies were carried out at varied concentrations of substrates in order to determine the true Michaelis constants and to elucidate the kinetic mechanism of reaction. Product-inhibition studies were carried out with each product (ADP, Pi, and oxalacetate) in combination with every substrate (ATP, bicarbonate, and pyruvate). From these kinetic studies and the existing knowledge on biotin-containing carboxylases, a mechanism was proposed for the action of pyruvate carboxylase which involves three independently active sites on the enzyme, one for each substrate. The interactions between the sites were visualized as being mediated by carboxybiotin formed on the enzyme. A steady-state rate equation was derived that satisfied kinetic results obtained.

Quantitative Effects of Oxygen Supply on Mycelia Growth and Extracellular Polysaccharide of Ganoderma tsugae

2012 ◽  
Vol 217-219 ◽  
pp. 975-978
Author(s):  
Nukrob Narkprasom ◽  
Tzou Chi Huang ◽  
Yuan Kuang Guu
Keyword(s):  
Mass Transfer ◽  
Secondary Metabolite ◽  
Submerged Fermentation ◽  
Oxygen Supply ◽  
Cell Mass ◽  
Extracellular Polysaccharide ◽  
Mycelia Growth ◽  
High Level ◽  
Dry Cell

The quantitative effects of oxygen supply in terms of shaking speed and medium volume on the production of mycelia and extracellular polysaccharide (EPS) using Ganoderma tsugae in submerged fermentation were investigated. Mycelia growth required the proper shaking speed at 134 rpm for breaking a larger pellet into several smaller pellets. Furthermore, high level of medium volume for consumption of abundant nutrient is needed for maximum mycelia growth. For EPS, a high agitation is needed to promote a good mass transfer for achieving high product concentrations recovery. The appropriate medium volume was found to be 150.4 mL for aiding to produce a secondary metabolite by promoting the mass transfer of substrates. The dry cell mass (DCM)-EPS diagram of G. tsugae was created from the both production equations which this diagram is an useful tool for submerge fermentation industry for decision about to control the both productions.

Evidence for microbial attenuation of particle flux in the Amundsen Gulf and Beaufort Sea: elevated hydrolytic enzyme activity on sinking aggregates

Polar Biology ◽  
2011 ◽  
Vol 34 (12) ◽  
pp. 2007-2023 ◽  
Author(s):  
Colleen T. E. Kellogg ◽  
Shelly D. Carpenter ◽  
Alisha A. Renfro ◽  
Amélie Sallon ◽  
Christine Michel ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Particle Flux ◽  
Hydrolytic Enzyme ◽  
Beaufort Sea ◽  
Amundsen Gulf

An in situ cell marker for clonal analysis of development of the extraembryonic endoderm in the mouse*

Development ◽  
1984 ◽  
Vol 80 (1) ◽  
pp. 251-288
Author(s):  
R. L. Gardner
Keyword(s):  
Enzyme Activity ◽  
Malic Enzyme ◽  
Cell Mass ◽  
Differential Staining ◽  
Wild Type ◽  
Primitive Endoderm ◽  
Extraembryonic Endoderm ◽  
Blue Tetrazolium ◽  
Malic Enzyme Activity ◽  
Parietal Endoderm

Conditions were found for staining whole mid-gestation capsular parietal endoderms and visceral yolk sacs for malic enzyme activity that gave excellent discrimination between wildtype (Mod-1+/Mod-1+) cells and mutant (Mod-ln/Mod-1n) cells that lack the cytoplasmic form of the enzyme. Reciprocal blastocyst injection experiments were undertaken in which single primitive endoderm cells of one genotype were transplanted into embryos of the other genotype. In addition, Mod-1+/Mod-1+ early inner cell mass (ICM) cells were injected into Mod-1n/Mod-1n blastocysts, either in groups of two or three singletons or as daughter cell pairs. A substantial proportion of the resulting conceptuses showed mosaic histochemical staining in the parietal endoderm, visceral yolk sac, or in both these membranes. Stained cells were invariably intimately intermixed with unstained cells in the mosaic parietal endoderms. In contrast, one or both of two distinct patterns of staining could be discerned in mosaic visceral yolk sacs. The first, a conspicuously ‘coherent’ pattern, was found to be due to endodermal chimaerism; the second, a more diffuse pattern, was attributable to chimaerism in the mesodermal layer of this membrane. The overall distribution of cells with donor staining characteristics resulting from primitive endoderm versus early ICM cell injections was consistent with findings in earlier experiments in which allozymes of glucosephosphate isomerase were used as markers. The conspicuous lack of phenotypically intermediate cells in predominantly stained areas of mosaic membranes suggested that the histochemical difference between Mod-1+/Mod-1+ and Mod-1n/Mod-ln genotypes was cell-autonomous. This conclusion was strengthened by the results of staining mixed in vitro cultures of parietal endoderm in which presence or absence of phagocytosed melanin granules was used as an independent means of distinguishing wild type from null cells. By substituting tetranitro blue tetrazolium for nitro blue tetrazolium in the incubation medium, satisfactory differential staining was obtained for both the extraembryonic endoderm and other tissues of earlier postimplantation wild type versus null embryos. Finally, absence of cytoplasmic malic enzyme activity does not appear to have a significant effect on the viability or behaviour of mutant cells.

scholarly journals PRODUCTION AND CHARACTERIZATION OF CELLULOLYTIC ENZYMES BY ASPERGILLUS NIGER AND RHIZOPUS SP. BY SOLID STATE FERMENTATION OF PRICKLY PEAR

2016 ◽  
Vol 29 (1) ◽  
pp. 222-233 ◽  
Author(s):  
TAMIRES CARVALHO DOS SANTOS ◽  
GEORGE ABREU FILHO ◽  
AILA RIANY DE BRITO ◽  
AURELIANO JOSÉ VIEIRA PIRES ◽  
RENATA CRISTINA FERREIRA BONOMO ◽  
...  
Keyword(s):  
Solid State ◽  
Aspergillus Niger ◽  
Water Activity ◽  
Solid State Fermentation ◽  
Cellulase Production ◽  
Cellulolytic Enzymes ◽  
Optimum Conditions ◽  
Thermostable Enzymes ◽  
Fermentation Time ◽  
Rhizopus Sp

ABSTRACT: Prickly palm cactus husk was used as a solid-state fermentation support substrate for the production of cellulolytic enzymes using Aspergillus niger and Rhizopus sp. A Box-Behnken design was used to evaluate the effects of water activity, fermentation time and temperature on endoglucanase and total cellulase production. Response Surface Methodology showed that optimum conditions for endoglucanase production were achieved at after 70.35 h of fermentation at 29.56°C and a water activity of 0.875 for Aspergillus niger and after 68.12 h at 30.41°C for Rhizopus sp. Optimum conditions for total cellulase production were achieved after 74.27 h of fermentation at 31.22°C for Aspergillus niger and after 72.48 h and 27.86°C for Rhizopus sp. Water activity had a significant effect on Aspergillus niger endoglucanase production only. In industrial applications, enzymatic characterization is important for optimizing variables such as temperature and pH. In this study we showed that endoglucanase and total cellulase had a high level of thermostability and pH stability in all the enzymatic extracts. Enzymatic deactivation kinetic experiments indicated that the enzymes remained active after the freezing of the crude extract. Based on the results, bioconversion of cactus is an excellent alternative for the production of thermostable enzymes.

Convenient Partial Purification of Glucose Oxidase from Aspergillus niger A9 Fermentation Broth by Reversed Micelles

2012 ◽  
Vol 554-556 ◽  
pp. 957-961
Author(s):  
Hong An ◽  
Xi Feng He ◽  
Shu Gang Gao
Keyword(s):  
Enzyme Activity ◽  
Aspergillus Niger ◽  
Glucose Oxidase ◽  
Fermentation Broth ◽  
Volume Ratio ◽  
Reversed Micelles ◽  
Partial Purification ◽  
Ammonium Bromide ◽  
Ultrasonic Oscillation ◽  
Protein Activity

Aim of this work was to establish the optimum conditions for the extraction and recovery by cationic reversed micelles of glucose oxidase (GOX) from Aspergillus niger A9, The influence of pH, temperature, solvent/co-solvents ratio on the extraction was investigated by experiment, using the residual enzyme activity to evaluate the results. The best condition for GOX extraction were ensured using iso-octane as solvent and butanol and n-hexanol co-solvent at 76/18/6 volume ratio, pH 4.80, 200mM cetyl-trimethyl ammonium bromide (CTAB) as cationic surfactant, The enzyme activity of GOX is measured by DNS method (3,5-dinitro salicylic acid method). In the extraction process, ultrasonic oscillation was adopted to mix organic solvent and water, ultrasonic oscillation temperature is 45 °C. Protein activity recovery of GOX can reach 88.2% in extraction.

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