scholarly journals ISOLASI DAN KARAKTERISASI EKSTRAK KASAR ENZIM BROMELIN DARI BATANG NANAS (Ananas comusus L.merr)

2006 ◽  
Vol 12 (1) ◽  
pp. 75-77
Author(s):  
Nuniek Herdyastuti

Brommelain is an enzyme hydrolyze most soluble protein easily and efficiently. This enzyme is used in many industry like food industry. This research aimed to isolation and characterization crude extract brommelain. This enzyme has been extracted from the stems of pineapples to produce crude extract, precipitated with amonium sulfat, and enzyme activity to decided with Bergmeyer methode. The higher activity was 1,996 U/ml in precipitate 40-60 percent amonium sulfat. Optimum temperature and pH to decided temperature and pH variation was detected based on enzyme activity. Characterization to indicate that bromelain has an optimum temperature at 55°C, optimum pH of 7, KM = 5.074 mg/ml and Vmax = 0.666 mg/ml.second.

2012 ◽  
Vol 457-458 ◽  
pp. 476-479
Author(s):  
Jing Xuan Gou ◽  
Wen Bin Dong ◽  
Qiao Zeng ◽  
Jing Jing Zhang

Chitosan is an aboundant biopolymer like cellose. To bioprocess them with chitin deacetylase (CDA), the products will be used in medicine and food industry. [In order to get new strains with significant ability of (CDA) Twenty eight strains producing CDA were isolated and screened out from the soil samples by color reaction in plate medium. The strain F2-7-3 was screened out from these strains for the highest CDA activity, which can reach more than 250U/mL. The paranitroacetanilide was used as the substrate, the optimum temperature for enzyme activity was determined to be 50°C and the optimum pH was 7.0. It was studied by method of morphological, physiological and biochemical characteristics and 16S rDNA analysis. Data shows that the strain was Rhodococcus sp.


2012 ◽  
Vol 12 (2) ◽  
pp. 124
Author(s):  
Zusfahair Zusfahair ◽  
Tien Setyaningtyas ◽  
Amin Fatoni

A bacterial lipase producer was isolated from garbage dump soil and was identified its genus. Lipase was extractedaccording to production time optimized, purified using ammonium sulfate fractionation and gel chromatograph.Determination of enzyme characteristic studied were influence of pH, temperature, various metals to lipaseactivity. The result of this research shows that the genus of isolated bacteria which produced lipase wasAcinetobacter sp., the lipase optimum production time is about 18 hours with the activity is about 115 unit/mL. Thehighest activity of lipase fractionation using ammonium sulfate is about 45% and the highest activity of purifyingwith filtration gel chromatograph column using Sephadex G-150 at 24 th fraction. Lipase from crude extract andpurifying product at this fraction has optimum pH 6 and optimum temperature is about 40 oC. Lipase to be classifiedas metalloenzyme that shows with decreasing the activity after added the EDTA. Metals ion, such as Cu 2+ and Zn2+were inhibited the lipase activity. Ca 2+ ion could increase lipase crude extract activity but inhibited the activity oflipase purifying product. Hg2+ ion could increase the activity of lipase purifying product.


2017 ◽  
Vol 2 (6) ◽  
pp. 172
Author(s):  
Emma Rochima ◽  
Rusky Intan Pratama ◽  
Yuli Andriani

The objective of this research was to isolate and characterize collagenase of Bacillus thuringiensis obtained from the collection of Aquatic Biotechnology Laboratory, Faculty of Fisheries and Marine Science, Padjadjaran University.  The substrate of collagen was derived from skin Tilapia waste of Cirata Reservoar. This study showed the presence of clear zone which is a sign of colagenolitik activity of B. thuringiensis. The optimum production time of collagenase was 24 hours of incubation. Collagenase of crude extract had collagenase activity of 0.181 units/ml with the protein concentration was 0.640 mg/ml. It was also found that the optimum temperature of collagenase derived from crude extract was 50° C and the optimum pH was 7-9. Keywords: collagenase; Bacillus thuringiensis;  skin, waste


Molekul ◽  
2009 ◽  
Vol 4 (2) ◽  
pp. 115
Author(s):  
Ari Asnani ◽  
Puji Lestari

The ability of Peryonix excavatus to live in extremely dirty area indicates that P. excavatus secretes distinctive enzymes which might be useful for industry. Thus, this research were aimed to isolate amylase, lipase and protease from P. excavatus, and to characterize the enzymes to know the optimum temperature and pH. The isolation procedure consisted of extraction and ammonium sulphate fractionation. The results showed that crude extract and ammonium sulphate fractions of P.excavatus had amylase, lipase, and protease enzymes activities. Among the three enzymes, amylase had the highest enzymatic activity whereas lipase was the least. The optimum temperature of amylase, lipase and protease were 60, 40, and 60 oC, respectively. The optimum pH of amylase, lipase and protease were 7, 7, and 8, respectively.


2017 ◽  
Vol 20 (2) ◽  
pp. 161
Author(s):  
Sri Wahyuni ◽  
R. Susanti ◽  
Retno Sri Iswari

This research aims to isolate and characterize the fcin enzyme from Ficus septica stem latex. Ficin from Ficus septica stem latex was isolated using column chromatography. Then enzyme activity was tested at different temperature (40oC, 50oC, 60oC, 70oC) and pH (6.0, 7.0, 8.0) levels. Ficin enzyme activity of joint treatment with variations in temperature and pH was analyzed using two-way ANOVA with a factorial pattern followed by Least Signifcant Difference (LSD) test. The results showed that temperature treatment signifcantly affects enzyme activity. However, the treatment of pH and the interaction between temperature and pH did not signifcantly affect the fcin enzyme activity. There was no signifcant difference in fcin activity at the incubation temperatures of 40oC and 50oC, as well as 60oC and 70oC. However, comparing the incubation temperatures of 40oC and 50oC with treatment 60°C and 70°C showed a signifcant difference in fcin enzyme activity. In the treatment of incubation at pH 6, 7 and 8 for fcin enzyme activity showed no signifcant difference. We concluded that the Ficus septica plant latex contained fcin enzyme with an optimum temperature of 60°C and optimum pH of 6, 7, and 8.


Molekul ◽  
2009 ◽  
Vol 4 (2) ◽  
pp. 73 ◽  
Author(s):  
Puji Lestari ◽  
Santi Nur Handayani ◽  
Oedjijono Oedjijono

Lipases are valuable biocatalysts because they act under extremely mild conditions, are stable in organic solvents, show broad substrate specificity and exhibit high stereoselectivity. Lipases play important role in various industries such as detergent, cosmetics, flavor, pharmacy and synthesis of organic compounds. The increasing of lipases requirements in industries is goading research to get new lipases resources commited. One of potential lipase resource is Azospirillum sp.JG3 bacteria from Microbiology Laboratory of Biology Faculty University of Jenderal Soedirman. The specific targets of this research are to get crude extract of lipase and investigate its biochemical characteristics. The method used were rejuvenation of Azospirillum sp.JG3 bacteria, inoculum production, determination of optimum production time and bacterium growth phase, extraction and production of lipase to get crude extract, and characterization the biochemical properties of lipase crude extract. The research resulted that crude extract of lipase from Azospirillum sp.JG3 had optimum temperature at 40 °C and optimum pH at pH 7. The lipase was a metalloenzyme with Ca2+ as its cofactor. The lipase was stable in three organic solvents tested, (chloroform, n-hexane and ether).


2016 ◽  
Vol 5 (1) ◽  
pp. 1
Author(s):  
Omar M. Atrooz

<p>β-galactosidase (EC 3.2.1.23) was extracted from <em>Cuminum cyminum </em>and<em> Curcuma longa</em>. The crude extracts of these plants were then characterized in term of pH, temperature, and enzyme kinetic. The crude extracts were also used in hydrolysis of lactose in milk and whey. The enzyme activity was measured by its ability to hydrolyze the substrate o-nitrophenyl β -D-galactopyranoside (ONPG).</p><p>It was found that β-galactosidase in the crude extracts of <em>Cuminum cyminum </em>exhibited maximum activity at pH 8.0 and optimum temperature at 60 °C. While, β-galactosidase in the crude extracts of <em>Curcuma longa</em> have optimum pH at 5.0 and 7.0 and optimum temperature at 50 °C.The K<sub>m</sub> and V<sub>max</sub> values of the β-galactosidase in the crude extracts of <em>Cuminum cyminum</em> and <em>Curcuma longa </em>were 4.16 mM and 0.087 μmol/min, and 2.63 mM and 0.333μmol/min, respectively.</p><p>The results showed that 96.84-97.08% of lactose was hydrolyzed in cow’s milk and whey when treated with crude extracts of <em>Cuminum cyminum</em> and 90-98.6% when treated with crude extracts of <em>Curcuma longa</em>.</p>


2019 ◽  
Vol 64 (4) ◽  
pp. 539-547
Author(s):  
Mohammed Inuwa Ja’afaru ◽  
Konjerimam Ishaku Chimbekujwo ◽  
Obinna Markraphael Ajunwa

Proteases are important industrial enzymes and fungi prove to be good sources of such enzymes. Purification techniques are however necessary for increased specificity in activity and better industrial value. Based on this, a protease produced by a Fusarium oxysporum was purified to homogeneity by Sephadex G-200 column and α–casein agarose chromatography. The enzyme had a molecular weight of 70 kDa in SDS-PAGE. Purified Fusarium oxysporum protease had a specific activity of 93.88 U/mg protein. The purification magnitude was 7.7 and the total yield was 20 %. Purified protease had an optimum pH of 5.0 while the optimum temperature was 40 °C. The enzyme was also thermotolerant (approximately 100 % at 40 °C for 2 h). The enzyme activity was stimulated by surfactants and metal ions like, Tween-20 and Mg2+. Enzyme activity was inhibited in presence of PMSF and EDTA. Casein was found to be the best substrate for protease activity of Fusarium oxysporum FWT1. Protease were tested upon blood stain for de-clotting of blood and was found to exhibit good de-clotting and de-staining activity after 15 minutes treatment time.


2012 ◽  
Vol 457-458 ◽  
pp. 472-475
Author(s):  
Jing Xuan Gou ◽  
Wen Bin Dong ◽  
Qiao Zeng ◽  
Lei Jin

Chitin is an abundant biopolymer like cellulose that is rather resistant to degradation. In order to develop a bio-digesting method, soil sample in Qinling Mountain were collected for screening the bacteria with high chitinase activity by method of the transparent circle. The strain D5-23 was isolated and screened out from soil, which was found with amazing chitinase acitivity. The ratio of transient circle and colony circle is no less than 10. The strain was then identified as Aeromonas sp according to the sequences of 16S rDNA and morphological analysis. The enzyme activity was studied further, ,data shows that the optimum temperature was 45°C, which is similar to other Aeromonas sp, wheras the optimum pH is 5 and 9, which is more similar to Vibrio alginolyticus TK-22.


2012 ◽  
Vol 14 (1) ◽  
pp. 114
Author(s):  
Ace Baehaki ◽  
Rinto Rinto ◽  
Agus Ramiadi

The aim of this study was to bacteria isolation and characterization proteases from water swamp. The water swamp samples collected fromIndralaya for microbial isolation. Three isolates showed proteolytic index >1.00. The optimum pH of extraceluller proteases from A6S3,A4S3 and A15S3 were 7.5; 8.0; 8.0, respectively. The optimum temperature of A6S3, A4S3 and A15S3 protease were 40; 50; 50 oC,respectively. Effect of metal ion (Fe, K, Mn, and Zn) shown Fe and K were inhibit protease A6S3, all metal ion were inhibit protease A4S3and K, Mn and Zn inhibit protease A15S3. Study on the effect of metals ion and spesific inhibitors indicated that all protease weremetaloprotease. Their moleculer weights were determined by using SDS-PAGE and zymogram technique for A6S3 isolate were 70, 88, 106and 121 kD respectively. Whereas for isolate A4S3 was 138 kD and isolate A15S3 was 131 kD.


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