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Journal ArticleDOI

Production of pectin lyase from Geobacillus pallidus p26, purification, characterization and fruit juice application

01 Apr 2014-Acta Chimica Slovenica (Sciendo)-Vol. 7, Iss: 1, pp 57-63

TL;DR: The purified pectin lyase enzyme was used for getting fruits juices and it was found that yields of fruits juices increased when they were compared with control.

AbstractA bacterial strain was isolated from Pasinler hot spring, Erzurum, Turkey. The purifi ed thermophilic isolate was identifi ed as Geobacillus pallidus P26 and used to produce extracellular pectin lyase (EC 4.2.2.10). Pectin lyase enzyme was purifi ed 34 fold by using DEAE-cellulose anion exchange column chromatography and characterized. Molecular weight of the enzyme was determined as 56 kDa by using Sephadex G-100 gel fi ltration chromatography. Purifi cation of enzyme was verifi ed by SDS-PAGE. The pH- and temperature optima of enzyme were determined (pH 9.0 and 60 oC, respectively). Pectin lyase was mostly stable at 50 oC for 24 hours. Its' activity decreased to 50 % for 24 h at 60 o C. K M and V max were calculated as 24.8 mg/mL and 2+ but not by Mg 2+ . The purifi ed pectin lyase enzyme was used for getting fruits juices. It was found that yields of fruits juices increased when they were compared with control.

Topics: Pectin lyase (66%)

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Citations
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Journal ArticleDOI
Abstract: Pectinase is an important group of industrial enzymes. Pectinase manufacturing occupies about 10% of the overall enzyme production world over. The aim of this study is to produce pectin lyase from Schizophyllum commune using the mosambi (sweet lime) fruit peels as substrate in solid state fermentation. The cultural parameters optimized through response surface methodology showed maximum pectin lyase production of 480.45 U/mL at initial medium pH of 6, incubation temperature of 35 °C, time period of 1 day, substrate concentration of 15 g and 3 mL of inoculum size. A purification fold of 3.08 with 355 U/mg specific activity and 4.16% yield was obtained after purification. Enzyme immobilization was done by entrapment with sodium alginate and adsorption with chitosan. Chitosan immobilized enzyme exhibited best thermal stability in the range of 45–55 °C and pH 8.0–9.0. Enzyme activity was stimulated in the presence of Ca2+ and Mg2+ while EDTA inhibited the enzyme activity. Chitosan immobilized pectin lyase was stable up to six cycles of reuse. The pH and thermal stability of S. commune pectin lyase makes it an important enzyme for industrial use. The results showed that pectin lyase produced from S. commune has significant potential for applications in the detergent and fruit juice industry. The enzyme produced from citrus agro waste via the proposed optimized biotechnological process can be explored for multiple industrial applications.

24 citations


01 Jan 2015
TL;DR: It was determined that FAMEs method failed in the identification of thermophilic bacteria at the species level whereas BOX-PCR technique was quite successful in exhibiting genetic differences among thermal isolates.
Abstract: A survey of isolation was practised by obtaining water samples from various thermal facilities in order to find out new types of thermophilic bacteria and to characterize them via phenotypic and genotypic methods. Hence, the test strains were first grouped according to their colonial morphologies and were characterized via conventional (morphological, physiological, biochemical) and molecular [Fatty Acid Methyl Ester profiles (FAME), 16S rRNA sequencing and BOX-PCR] methods. As the result of the research, it was determined that FAMEs method failed in the identification of thermophilic bacteria at the species level whereas BOX-PCR technique was quite successful in exhibiting genetic differences among thermal isolates. On the other hand, as the result of 16S rRNA sequence, 4 of the isolated strains (A7-A10) were determined to resemble Bacillus licheniformis at a rate of ≥97% while 2 of them (A1 and A13) resembled to Anoxybacillus kaynarcensis, 1 of them (A2) to A. flavithermus, 1 of them (A3) to A. rupiensis, 1 of them (A4) to A. thermarum, 1 of them (A5) to A. gonensis, 1 of them (A11) to B. thermoamylovorans,1 of them (A12) to Ureibacillus thermospaericus, 1 of them (A14) to Brevibacillus brevis, 1 of them (A6) to B. thermolactis and again 1 of them resembled to B. thermoruber (Z1) at the same rate. Their amylase and cellulase productions were also examined and analyzed and the enzyme producing isolates that were biotechnologically valuable were presented to be used in the prospective studies.

12 citations



Journal ArticleDOI
TL;DR: A new potent strain of thermophilic bacterium isolated from Sungai Klah Hot Spring Park in Perak, Malaysia for the first time is revealed and the high production of thermostable protease enzyme by G. thermoglucosidasius SKF4 highlighted the promising properties of this bacterium for industrial and biotechnological applications.
Abstract: Major progress in the fields of agriculture, industry, and biotechnology over the years has influenced the quest for a potent microorganism with favorable properties to be used in scientific research and industry. This study intended to isolate a new thermophilic-protease-producing bacterium and evaluate its growth and protease production under cultural conditions. Protease producing bacteria were successfully isolated from Sungai Klah Hot Spring Park in Perak, Malaysia, and coded as SKF4; they were promising protease producers. Based on microscopic, morphological, and 16S rRNA gene analysis, isolate SKF4 was identified as Geobacillus thermoglucosidasius SKF4. The process of isolating SKF4 to grow and produce proteases under different cultural conditions, including temperature, pH, NaCl concentration, carbon and nitrogen sources, and incubation time, was explored. The optimum cultural conditions observed for growth and protease production were at 60 to 65 °C of temperature, pH 7 to 8, and under 1% NaCl concentration. Further, the use of casein and yeast extract as the nitrogen sources, and sucrose and fructose as the carbon sources enhanced the growth and protease production of isolate SKF4. Meanwhile, isolate SKF4 reached maximum growth and protease production at 24 h of incubation time. The results of this study revealed a new potent strain of thermophilic bacterium isolated from Sungai Klah Hot Spring Park in Perak, Malaysia for the first time. The high production of thermostable protease enzyme by G. thermoglucosidasius SKF4 highlighted the promising properties of this bacterium for industrial and biotechnological applications.

9 citations


Cites background from "Production of pectin lyase from Geo..."

  • ...from other studies has been also reported having the ability to grow within the range of NaCl concentration 0% to 5% [29,55]....

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Journal ArticleDOI
TL;DR: A thermophilic bacterium (TP-2) was isolated from the Tatta Pani hot spring in Azad Kashmir and was characterized using phenotypic and genotypic characters and Sequence analysis of its 16S rRNA gene showed that isolate TP-2 had 89% homology with Geobacillus debilis.
Abstract: A thermophilic bacterium (TP-2) was isolated from the Tatta Pani hot spring in Azad Kashmir and was characterized using phenotypic and genotypic characters. The strain developed cream colored, round, smooth, flat and slimy colonies while the cells were Gram positive rods that ranged in size from about 2.1-3.6 μm to 0.2-0.3 μm in width. Sequence analysis of its 16S rRNA gene showed that isolate TP-2 had 89% homology with Geobacillus debilis. It grew within pH range of 5.5 to 8.5 with optimum growth at pH 7.0. The isolate showed optimum growth at 65oC and gave positive results for gelatin hydrolysis (GEL), ortho nitrophenyl-β-D-galactopyranosidase (ONPG), and nitrate production and produced acid from sucrose, glucose and maltose. It utilized glucose, fructose, maltose, lactose, sucrose, xylan, starch, filter paper and carboxymethylcellulose as sole carbon source. Isolate TP-2 produced significant amount of industrially important enzymes i.e. extracellular α-amylase, CMCase, FPase, Xylanase, Protease and Lipase and intracellular CMCase and FPase.

8 citations


Cites background from "Production of pectin lyase from Geo..."

  • ...Geobacillus pallidus p26 grew within the NaCl range of 2-5% (Celik et al. 2014)....

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  • ...Geobacillus pallidus p26 isolated from Pasinler hot spring, Erzurum, Turkey grew optimally at pH 7.5 (Celik et al. 2014)....

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References
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Journal ArticleDOI
Abstract: Pectinases or petinolytic enzymes, hydrolyze pectic substances. They have a share of 25% in the global sales of food enzymes. Pectinases are one of the most widely distributed enzymes in bacteria, fungi and plants. Protopectinases, polygalacturonases, lyases and pectin esterases are among the extensively studied pectinolytic enzymes. Protopectinases catalyze the solubilization of protopectin. Polygalacturonases hydrolyze the polygalacturonic acid chain by addition of water and are the most abundant among all the pectinolytic enzymes. Lyases catalyze the trans-eliminative cleavage of the galacturonic acid polymer. Pectinesterases liberate pectins and methanol by de-esterifying the methyl ester linkages of the pectin backbone. Pectinolytic enzymes are of significant importance in the current biotechnological era with their all-embracing applications in fruit juice extraction and its clarification, scouring of cotton, degumming of plant fibers, waste water treatment, vegetable oil extraction, tea and coffee fermentations, bleaching of paper, in poultry feed additives and in the alcoholic beverages and food industries. The present review mainly contemplates on the types and structure of pectic substances, the classification of pectinolytic enzymes, their assay methods, physicochemical and biological properties and a bird's eye view of their industrial applications.

883 citations


"Production of pectin lyase from Geo..." refers background in this paper

  • ...…those are fruit juice processing, coffee and tea processing, macerating of plants and vegetable tissue, degumming of plant fibers, treatment waste water, extracting vegetable oil, bleaching of paper, adding poultry feed and the textile, alcoholic beverages, and food industries (Jayani et al. 2005)....

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Book
01 Aug 1989

401 citations


Journal ArticleDOI
TL;DR: The idea is now emerging that this type of yeast enzyme could offer an alternative to fungal enzymes for industrial applications.
Abstract: When grown in the appropriate medium, several yeast species produce pectinases able to degrade pectic substances. It is mainly exocellular endopolygalacturonases that break pectins or pectate down by hydrolysis of α-1,4-glycosidic linkages in a random way. Biochemical characterisation of these enzymes has shown that they have an optimal pH in the acidic region and an optimal temperature between 40 and 55°C. Their production by yeasts is a constitutive feature and is repressed by the glucose concentration and aeration. Pectic substances and their hydrolysis products are used as carbon sources by a limited number of yeasts and hence these enzymes must be involved in the colonisation of different parts of plants, including fruits. The first yeast pectic enzyme (encoded by the PSE3 gene) was cloned from Tichosporon penicillatum. Recently, a polygalacturonase-encoding gene from Saccharomyces cerevisiae has been cloned and overexpressed in several strains and the gene for an extracellualar endopolygalacturonase from Kluyveromyces marxianus has also been described. Taking all the results together, the idea is now emerging that this type of yeast enzyme could offer an alternative to fungal enzymes for industrial applications.

211 citations


"Production of pectin lyase from Geo..." refers background in this paper

  • ...Pectinases are produced by a large number of organisms, such as bacteria (Kim et al. 1998), fungi (Alana et al. 1991, Obi et al. 1985), actinomycetes (Beg et al. 2000) and yeast (Blancoa et al. 1999)....

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