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.

Pectinase Production from Schizophyllum commune Through Central Composite Design Using Citrus Waste and Its Immobilization for Industrial Exploitation

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.

https://www.mdpi.com/1420-3049/25/11/2609/pdf

Effect of Cultural Conditions on Protease Production by a Thermophilic Geobacillus thermoglucosidasius SKF4 Isolated from Sungai Klah Hot Spring Park, Malaysia.

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Geobacillus Bacteria: Potential Commercial Applications in Industry, Bioremediation, and Bioenergy Production

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.

Identification of Thermophilic Strains from Geothermal Areas in Turkey by using Conventional and Molecular Techniques

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.

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Characterization Of A Novel Hydrolytic Enzyme Producing Thermophilic Bacterium Isolated From The Hot Spring Of Azad Kashmir-Pakistan

A 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.

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

Hipertansiyon, cogu yasli insanin mustarip oldugu bir hastaliktir. Yuksek tansiyonu olan insanlar genellikle hayatlari boyunca saglikli bir yasam surdurebilmek icin kan basincini dusuren ilaclari kullanmak zorundadirlar. Bu calismada, lisiniprol adli hipertansiyon ilacinin kemik yapim ve yikiminin bir gostergesi olan alkalen fosfataz (ALP) ve tartarata dayanikli asit fosfataz (TrACP) enzimlerinin aktiviteleri uzerine olan etkilerini in vitro olarak incelenmistir. Insan serumundan alkalen fosfataz enzimi DEAE-seluloz, asit fosfataz enzimi ise CM-seluloz iyon degisim kromatografisi kullanilarak saflastirilmistir. Alkalen ve asit fosfataz enzimlerinin aktiviteleri substrat olarak p-nitrofenilfosfat kullanilarak belirlenmistir. TrACP enzim aktivitesi bulunurken ortama tartarat ilave edilerek TrACP disindaki diger asit fosfataz enzimlerinin inhibe olmasi saglanmistir. Alkalen fosfataz ve tartarata dayanikli asit fosfataz enzimlerinin aktiviteleri substratin (0,033, 0,1, 0,16, 0,23 ve 0,3 mM) bes farkli konsantrasyonunda inhibitorlu ve inhibitorsuz ortamlarda bulunmustur. Inhibitorun ise her bir substrat konsantrasyonu icin 10 -2 , 10 -3 ve 10 -4 M olmak uzere uc farkli derisimi kullanilmistir. Ilacin her iki enzimi de inhibe ettigi tespit edilip, I 50 degerleri 0,17 mM  ve 0,79 mM olarak hesaplanmistir. Ayrica K i degerleri Lineweaver-Burk grafiginden yararlanilarak ALP ve TrACP icin sirasiyla 6,98x10 -4 , 7,06x10 -4 olarak hesaplanmistir. Hypertension is a disease that most elderly people suffer. People with high blood pressure often obliged to use the drug that lowers blood pressure in order to maintain a healthy lifestyle throughout their lives. In this study, the in vitro effect of lisonopril on serum alkaline phosphatase and acid phosphatase enzymes which are essential for normal bone formation and mineralization activities was investigated. Alkaline phosphatase enzyme was purified using DEAE-cellulose, acid phosphatase enzyme was purified using CM-cellulose ion exchange chromatography from human serum. The activities of alkaline and acid phosphatase were determined using p-nitrophenylphosphate as the substrate. When finding out the activity of TrACP enzyme, adding tartrate to the environment provided inhibition of other acid phosphatase enzymes than TrACP. The activities of alkaline phosphatase and acid phosphatase, which is resistant to tartrate, have been found at five different concentrations of substrates (0.033, 0.1, 0.16, 0.23 and 0.3 mm) in environment with inhibitor and without inhibitor. While three different concentrations of the inhibitor, including 10 -2 , 10 -3 and 10 -4 , was used for each substrate concentration. It was found that the drug inhibit both enzymes, and I 50 value was calculated as 0.17 mm and 0.79 mm. Furthermore, Ki values ​​were calculated as 6.98x10 -4 , 7.06x10 -4 for ALP and TrACP respectively by utilizing the Lineweaver-Burk plot.

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Serum Alkalen Fosfataz ve Asit Fosfataz Enzimlerinin Aktivileri Üzerine Lizinoprilin İn Vitro Etkisi / The in vitro effect of lisonopril on serum alkaline phosphatase and acid phosphatase enzymes activity

Safinur Yıldırım Çelik

Papers

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

Serum Alkalen Fosfataz ve Asit Fosfataz Enzimlerinin Aktivileri Üzerine Lizinoprilin İn Vitro Etkisi / The in vitro effect of lisonopril on serum alkaline phosphatase and acid phosphatase enzymes activity