scholarly journals Purification of an Exopolygalacturonase fromPenicillium viridicatum RFC3Produced in Submerged Fermentation

2009 ◽  
Vol 2009 ◽  
pp. 1-8 ◽  
Author(s):  
Eleni Gomes ◽  
Rodrigo Simões Ribeiro Leite ◽  
Roberto da Silva ◽  
Dênis Silva
Keyword(s):  
Molecular Weight ◽  
Submerged Fermentation ◽  
Galacturonic Acid ◽  
Apparent Molecular Weight ◽  
Degree Of Esterification ◽  
Optimum Ph ◽  
The Stability ◽  
High Degree ◽  
Hydrolysis Of ◽  
Optimum Ph And Temperature

An exo-PG obtained fromPenicillium viridicatumin submerged fermentation was purified to homogeneity. The apparent molecular weight of the enzyme was 92 kDa, optimum pH and temperature for activity were pH 5 and 50–55∘C. The exo-PG showed a profile of an exo-polygalacturonase, releasing galacturonic acid by hydrolysis of pectin with a high degree of esterification (D.E.). IonsCa2+enhanced the stability of enzyme and its activity by 30%. TheKmwas 1.30 in absence ofCa2+and 1.16 mgmL−1in presence of this ion. In relation to theVmaxthe presence of this ion increased from 1.76 to 2.07 μmolmin−1mg−1.

Characterisation of a Neutral Protease Produced by Micrococcus luteus

1996 ◽  
Vol 51 (5-6) ◽  
pp. 429-431 ◽  
Author(s):  
M.O. Ilori ◽  
O.O. Amund ◽  
O. Omidiji
Keyword(s):  
Molecular Weight ◽  
Citric Acid ◽  
Proteolytic Enzyme ◽  
Micrococcus Luteus ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Optimum Temperature ◽  
Optimum Ph

Abstract A proteolytic enzyme produced by a cassava-ferment­ing strain of Micrococcus luteus was extracted and puri­fied 50-fold by gel filtration and ion exchange chromatography. The optimum pH for the enzyme was 7.0, the optimum temperature 25 °C, the apparent molecular weight 42 kDa and the Km value, 0.45 mg ml-1 with casein as substrate. The enzyme was stimulated by Ca2+ and Mg2+ but inhibited by Zn2+ and Co2+ ions. Other inhibitors were EDTA, KCN, citric acid and L-cysteine indicating the enzyme to be a metalloprotease.

scholarly journals Isolation of Corncob Xylan-Degrading Fungi and Its Application in Xylobiose Production

2021 ◽  
Vol 20 (2) ◽  
Author(s):  
Tantry Febrinasari ◽  
Hasegawa Tae ◽  
Nakanishi Riki ◽  
Akkharapimon Yotsombat ◽  
Takata Goro ◽  
...  
Keyword(s):  
Particle Size ◽  
Incubation Temperature ◽  
Xylanase Activity ◽  
Solid Substrate ◽  
Enzyme Loading ◽  
Content Ratio ◽  
Optimum Ph ◽  
Initial Ph ◽  
Hydrolysis Of ◽  
Optimum Ph And Temperature

In the present study, a potential corncob xylan degradation fungi was isolated and screened from soil to produce xylanase, and was identified as Fusarium oxysporum. The production of xylanase by F. oxysporum under solid state fermentation using corncob powder as the solid substrate reached the maximum xylanase activity when using particle size of substrate of 60 mesh, water content ratio of 2 mL/g substrate, incubation temperature of 30°C, initial pH of 6.0, size of inoculum of 5x107 spore/3 g substrate, and incubation time of 2 days. The xylanase activity increased about 4 times up to 7.92 U/mL after optimization. The potential application of xylanase of F. oxysporum in hydrolyzing alkali-treated corncob xylan to produce xylobiose was also demonstrated. Hydrolysis of 6% of corncob xylan using 100 U/g substrate of enzyme loading under optimum pH and temperature conditions (pH 5.5 and 50°C, respectively) achieved the yield of xylobiose up to 28.7 g/100 g pure xylan after 12 h incubation. The purification of hydrolysate could retain 91.1% of xylobiose. Further separation step using activated charcoal column chromatography was able to get a pure xylobiose, but could only recover 59.3% of xylobiose.

scholarly journals Production and Characterization of Crude Protease from RS1 Isolate from silage of Floating Bladderwort (Utricularia gibba)

2020 ◽  
Vol 10 (3) ◽  
pp. 289-293
Author(s):  
Ace Baehaki ◽  
Arif Hidayat ◽  
Nuni Gofar ◽  
Rodiana Nopianti
Keyword(s):  
Molecular Weight ◽  
Production Time ◽  
Molecular Weights ◽  
Sds Page ◽  
Optimum Ph ◽  
Utricularia Gibba ◽  
Optimum Production ◽  
Optimum Ph And Temperature

The purpose of this research was to produce and characterizing crude protease from RS1 isolate of swamp plant silage. The optimum production time of RS1 isolate was 40 h. The optimum pH and temperature of protease from RS1 isolate were 10 and 45℃, respectively.  Ion Mg3+ increased RS1 protease whereas ion of Na+, K+, Fe2+, and Zn2+ inhibited protease from RS1 isolate. Study on the effect of metals ion indicated that protease from RS1 isolate was metaloenzyme. Based analysis on SDS-PAGE, the molecular weight of RS1 protease had 12 bands with molecular weights ranging from 34.75 kDa to 263.53 kDa.

Isolation and partial characterization of a peptidase with trypsin-like activity from bovine adenohypophysis

1989 ◽  
Vol 54 (8) ◽  
pp. 2276-2286
Author(s):  
Tsezengijn Dash ◽  
Tomislav Barth ◽  
Jiřina Slaninová ◽  
Jana Barthová ◽  
Hana P. Mašková ◽  
...  
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Partial Characterization ◽  
Chromogenic Substrate ◽  
Fluorogenic Substrate ◽  
Basic Amino Acids ◽  
Optimum Ph ◽  
Reproducible Method ◽  
Hydrolysis Of

A reproducible method has been developed for the isolation of the adenohypophyseal enzyme with a trypsin-like activity. The enzyme is able to hydrolyze Nα-benzoyl-L-arginine-p-nitroanilide, a fluorogenic substrate CBzl-Arg-Arg-β-naphthyl amide and some peptides with one or two accumulated basic amino acids in the chain. The optimum pH for hydrolysis of the chromogenic substrate was within the range 6.0-7.0 (Km = 0.66 mmol l-1), in the case of the fluorogenic substrate the range was between 7.0 and 7.5 (Km = 1.2 μmol l-1). The enzyme is activated by cysteine and dithiothreitol and inhibited by SH-poisons. The molecular weight of the enzyme, determined by means of two independent methods, was approximately 25 kDA.

scholarly journals Molybdenum Reduction to Molybdenum Blue inSerratiasp. Strain DRY5 Is Catalyzed by a Novel Molybdenum-Reducing Enzyme

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
M. Y. Shukor ◽  
M. I. E. Halmi ◽  
M. F. A. Rahman ◽  
N. A. Shamaan ◽  
M. A. Syed
Keyword(s):  
Molecular Weight ◽  
Isoelectric Point ◽  
Catalytic Efficiency ◽  
Apparent Molecular Weight ◽  
Underlying Mechanism ◽  
Maximum Activity ◽  
Molybdenum Blue ◽  
Optimum Ph ◽  
Substrate Concentrations

The first purification of the Mo-reducing enzyme fromSerratiasp. strain DRY5 that is responsible for molybdenum reduction to molybdenum blue in the bacterium is reported. The monomeric enzyme has an apparent molecular weight of 105 kDalton. The isoelectric point of this enzyme was 7.55. The enzyme has an optimum pH of 6.0 and maximum activity between 25 and 35°C. The Mo-reducing enzyme was extremely sensitive to temperatures above 50°C (between 54 and 70°C). A plot of initial rates against substrate concentrations at 15 mM 12-MP registered aVmaxfor NADH at 12.0 nmole Mo blue/min/mg protein. The apparentKmfor NADH was 0.79 mM. At 5 mM NADH, the apparentVmaxand apparentKmvalues for 12-MP of 12.05 nmole/min/mg protein and 3.87 mM, respectively, were obtained. The catalytic efficiency (kcat/Km) of the Mo-reducing enzyme was 5.47 M-1 s-1. The purification of this enzyme could probably help to solve the phenomenon of molybdenum reduction to molybdenum blue first reported in 1896 and would be useful for the understanding of the underlying mechanism in molybdenum bioremediation involving bioreduction.

scholarly journals Performance Comparison of α- and β-Amylases on Chitosan Hydrolysis

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Nur Rokhati ◽  
Prita Widjajanti ◽  
Bambang Pramudono ◽  
Heru Susanto
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
High Viscosity ◽  
Performance Comparison ◽  
Optimum Ph ◽  
Hydrolysis Temperature ◽  
Hydrolysis Of ◽  
Low Solubility ◽  
Chitosan Hydrolysis ◽  
A Performance

The low solubility in common solvent and high viscosity resulting from its high molecular weight (MW) with fiber-like structure prevents a more widespread use of chitosan. This paper presents a performance comparison of nonspecific, commercially available enzymes, α- and β-amylases, for the hydrolysis of chitosan to lower its MW. The results showed that both enzymes demonstrate the ability to be used as catalysts in chitosan hydrolysis with β-amylase having better performance than α-amylase. The chitosan hydrolysis was influenced by not only the enzyme and the chitosan characteristics but also the hydrolysis condition. The optimum pH solution was 4 for α-amylase and 5 for β-amylase. The hydrolysis temperature was found to be optimal at 90 and 50°C for α- and β-amylases, respectively.

scholarly journals Isolasi Mikroorganisme Potensial Penghasil Lipase dari Limbah Pengolahan Minyak Kelapa Sawit Malinping

2020 ◽  
Vol 13 (2) ◽  
pp. 228-241
Author(s):  
Ika Rahmatul Layly ◽  
Erma Widyasti ◽  
Deden Rosid Waltam ◽  
Ayi Mufti ◽  
Nita Wiguna ◽  
...  
Keyword(s):  
Crude Oil ◽  
Fats And Oils ◽  
Optimum Temperature ◽  
Processing Waste ◽  
Titration Method ◽  
Optimum Ph ◽  
Oil Tank ◽  
Textile Industries ◽  
Hydrolysis Of ◽  
Optimum Ph And Temperature

AbstrakLipase adalah kelompok enzim yang mengkatalisis hidrolisis rantai panjang trigliserida, lemak, dan minyak menjadi gliserol dan asam lemak dengan adanya air. Sumber lipase untuk industri kebanyakan berasal dari mikroorganisme. Penggunaan lipase pada industri makin meningkat setiap tahunnya meliputi aplikasinya pada industri makanan, pakan, farmasi, pulp, dan kertas, biodiesel, dan industri tekstil. Dalam usaha mendapatkan isolat potensial penghasil lipase untuk hHidrofilisasi serat poliester, pada penelitian ini dilakukan skrining dan isolasi mikroorganisme yang dapat menghasilkan lipase dari limbah pengolahan minyak kelapa sawit di Malinping, Lebak, Banten. Sebanyak 20 isolat bakteri dan 5 isolat jamur yang diperoleh kemudian diuji aktivitas lipasenya menggunakan metode titrasi. Empat isolat bakteri terpilih (Kondensat, Lumpur-Got, Hasil-Buangan, dan Tangki-Crude-Oil) serta lima isolat jamur (Nut-A, Nut-B, Nut-C, Kernel-B, dan Kernel-C) dikarakterisasi pH dan suhu optimum enzimnya. Hasil karakterisasi pH menunjukkan bahwa isolat bakteri Kondensat, Lumpur-Got, Hasil-Buangan, dan Tangki-Crude-Oil mempunyai aktivitas enzim lipase tertinggi pada pH 6. Suhu optimal aktivitas enzim lipase isolat Lumpur-Got-B, Hasil Buangan-B, dan Tangki-Crude-Oil B  pada 40 °°C, sedangkan isolat bakteri-Kondensat-B optimal pada suhu 30 °°C. Aktivitas lipase kelima isolat jamur optimal pada pH 6. Suhu optimal aktivitas lipase isolat jamur Nut-A adalah 40 °°C, sedangkan isolat Nut-B, Nut-C, Kernel-B, dan Kernel-C aktivitasnya optimal pada 50 °°C.Abstract Lipase are enzymes that catalyzed the hydrolysis of triglyceride, fats and oils into glycerol and fatty acids in the presence of water. Industrial Lipase source mostly derived from microbes. Each year, the lipase utilization in industry increased, such as application for foods, feeds, pharmacys, pulp and papers, biodiesel, and textile industries. On this study, a total of 20 bacteria and 5 fungi lipase potential producer were screened and isolated from oil palm processing waste in Malinping, Lebak, Banten, which then tested for its activity using titration method. Selected isolates then were characterized for its enzyme optimum pH and temperature. The optimum pH for isolate Kondensat, Lumpur-Got, Hasil-Buangan and Crude-Oil-Tank lipases are at pH 6, whilst the optimum temperature of isolates Lumpur-Got B, Hasil-Buangan B and Crude-Oil-Tank B were at 40 °°C and bakteri-Kondensat B isolate optimum at 30 °°C. The five fungi characterization shown optimum pH at 6 and 50 °°C except for isolate Nut-A that optimum at 30 °°C.

scholarly journals Phytase Expressed by pIAβ8 and pGAPZαA Vectors and Analysis of its Biochemical Characters

2009 ◽  
Vol 3 (1) ◽  
pp. 19-23 ◽  
Author(s):  
H. Feng ◽  
R.Y. Zuo ◽  
J. Chang ◽  
Q.H. Zheng ◽  
Q.Q. Yin
Keyword(s):  
Molecular Weight ◽  
Solid Medium ◽  
Expression Vectors ◽  
Aspergillus Ficuum ◽  
Coding Region ◽  
Phytase Gene ◽  
E Coli ◽  
Extracellular Phytase ◽  
Optimum Ph ◽  
Optimum Ph And Temperature

Phytase and phytase gene from Aspergillus ficuum (A. ficuum) were used in this study. The results showed that phytase activity reached the peak of 0.17 U/g after 4 d incubation in solid medium for A. ficuum; the optimum pH and temperature of phytase were 2.5 and 50 oC, respectively. A 1.4-kb DNA containing the coding region of phytase gene was isolated and inserted into the expression vectors of pIAβ8 and pGAPZαA, which were transformed into E. coli (Top 10). The maximal phytase activities in the supernatant and cells were 2.31 and 9.04 U/ml for the E. coli with pIAβ8, 8.04 and 2.93 U/ml for the E. coli with pGAPZαA, respectively. It was concluded that the recombinant of pIAβ8-phytase could express intracellular phytase, while the recombinant of pGAPZαA-phytase could express extracellular phytase. The molecular weight of phytase protein was 54.61 kDa.

scholarly journals A Comparative Study of New Aspergillus Strains for Proteolytic Enzymes Production by Solid State Fermentation

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Gastón Ezequiel Ortiz ◽  
Diego Gabriel Noseda ◽  
María Clara Ponce Mora ◽  
Matías Nicolás Recupero ◽  
Martín Blasco ◽  
...  
Keyword(s):  
Solid State ◽  
Comparative Study ◽  
Solid State Fermentation ◽  
Antioxidant Activities ◽  
Proteolytic Enzymes ◽  
Thermodynamics And Kinetics ◽  
Wide Range ◽  
Optimum Ph ◽  
Hydrolysis Of ◽  
Optimum Ph And Temperature

A comparative study of the proteolytic enzymes production using twelve Aspergillus strains previously unused for this purpose was performed by solid state fermentation. A semiquantitative and quantitative evaluation of proteolytic activity were carried out using crude enzymatic extracts obtained from the fermentation cultures, finding seven strains with high and intermediate level of protease activity. Biochemical, thermodynamics, and kinetics features such as optimum pH and temperature values, thermal stability, activation energy (Ea), quotient energy (Q10), Km, and Vmax were studied in four enzymatic extracts from the selected strains that showed the highest productivity. Additionally, these strains were evaluated by zymogram analysis obtaining protease profiles with a wide range of molecular weight for each sample. From these four strains with the highest productivity, the proteolytic extract of A. sojae ATCC 20235 was shown to be an appropriate biocatalyst for hydrolysis of casein and gelatin substrates, increasing its antioxidant activities in 35% and 125%, respectively.

scholarly journals Characterization of the purified Chlamydomonas minus agglutinin.

1985 ◽  
Vol 101 (3) ◽  
pp. 1144-1152 ◽  
Author(s):  
P Collin-Osdoby ◽  
W S Adair
Keyword(s):  
Molecular Weight ◽  
Apparent Molecular Weight ◽  
Agarose Beads ◽  
Sds Page ◽  
Sexual Agglutinins ◽  
Molecular Morphology ◽  
High Degree

Chlamydomonas flagellar sexual agglutinins are responsible for the adhesion of opposite mating-type (plus and minus) gametes during the first stages of mating. Purification and partial characterization of the plus agglutinin was previously reported (Adair, W. S., C. J. Hwang, and U. W. Goodenough, 1983, Cell, 33:183-193). Here we characterize the purified minus molecule. We show it to be a high molecular weight, hydroxyproline-rich glycoprotein that migrates in the 3% stacking region of an SDS-polyacrylamide gel and is absent from two nonagglutinating minus mutants. Plus and minus agglutinins are remarkably similar, although nonidentical, in amino acid composition, molecular morphology, and reactivity in vivo and in vitro with monoclonal antibodies raised against the plus agglutinin. Moreover, the adhesiveness of both plus and minus agglutinins, when coupled to agarose beads, is abolished by thermolysin, trypsin, periodate, alkaline borohydride, reducing agents, or heat, but unaffected by exo- or endoglycosidases. The minus agglutinin, however, migrates just ahead of the plus molecule on SDS PAGE, is excluded from an anion-exchange (Mono Q) column, elutes earlier during hydrophobic interaction (Bio-gel TSK Phenyl 5PW) chromatography, and is sensitive to chymotrypsin digestion (unlike the plus agglutinin); therefore, it differs from the plus agglutinin in apparent molecular weight, net charge, relative hydrophobicity and proteolytic susceptibility. Nevertheless, our results generally demonstrate a high degree of homology between these complementary cell-cell recognition/adhesion molecules, which suggests that they are specified by genes that have a common evolutionary origin.

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