Penicillium chrysogenum glucose oxidase - a study on its antifungal effects (original) (raw)
Related papers
Functional Foods in Health and Disease, 2023
Backgrounds: Food enzymes play a crucial role in enhancing specific food attributes, such as improving texture, eliminating toxins and allergens, producing carbohydrates, and enhancing flavor and appearance characteristics. Glucose oxidase (GOX) catalyzes the oxidation of glucose to produce gluconic acid and hydrogen peroxide, making it applicable in various practical scenarios. This enzyme could be utilized to eliminate glucose, thereby facilitating the creation of functional food suitable for individuals with diabetes and glucose intolerance. Our main goal was to characterize GOX from a Penicillium chrysogenum strain isolated from Armenian ecosystems. Methods: GOX of P. chrysogenum MDC 8358 was obtained by surface fermentation using glucose as a carbon source. The enzyme was purified 230-fold to homogeneity using a three-step purification scheme consisting of ion exchange, adsorption, and gel filtration chromatography. Results: The specific activity of the purified enzyme reached 271.2 U/mg. GOX from P. chrysogenum MDC 8358 has shown to be dimeric with a molecular weight of 135.5 kDa, consisting of two equal subunits with a molecular weight of 70 kDa. Isoelectric focusing has revealed a pH of 4.6. The enzyme has displayed a temperature optimum of 40 °C, and a pH optimum of 6.8, being more thermostable at acidic pH. Characterized GOX was highly specific for β-D-glucose and has shown only minor activities to mannose, xylose, and galactose. Keywords: Penicillium chrysogenum MDC 8358, surface fermentation, glucose oxidase, purification, characterization, functional food preservation
2000
A novel glucose oxidase (GOX), a Xavoenzyme, from Penicillium sp. was isolated, puriWed and partially characterised. Maximum activities of 1.08 U mg ¡1 dry weight intracellular and 6.9 U ml ¡1 extracellular GOX were obtained. Isoelectric focussing revealed two isoenzymes present in both intra-and extracellular fractions, having pI's of 4.30 and 4.67. GOX from Penicillium sp. was shown to be dimeric with a molecular weight of 148 kDa, consisting of two equal subunits with molecular weight of 70 kDa. The enzyme displayed a temperature optimum between 25 and 30°C, and an optimum pH range of 6-8 for the oxidation of-D-glucose. The enzyme was stable at 25°C for a minimum of 10 h, with a half-life of approximately 30 min at 37°C without any prior stabilisation. The lyophilized enzyme was stable at ¡20°C for a minimum of 6 months. GOX from Penicillium sp. Tt42 displayed the following kinetic characteristics: V max , 240.5 U mg ¡1 ; K m , 18.4 mM; k cat , 741 s ¡1 and k cat /K m , 40 s ¡1 mM ¡1. Stability at room temperature, good shelf-life without stabilisation and the neutral range for the pH optimum of this GOX contribute to its usefulness in current GOX-based biosensor applications.
Transformation and expression of Penicillium funicolusum glucose oxidase gene in yeast
2013
Glucose oxidase is an important enzyme hydrolyzing for its hydrolyzing activity on glucos. It possesses and has a wide board of applications in different industries such as bakery, pharmaceutical, plant pathology and biosensors. In this study, yeast (Saccharomyces cerevisiae) was transformed successfully by the glucose oxidase gene (gox) obtained from Penicillium funicolusum. The secreted glucose oxidase enzyme (GOX) by yeast transformants was characterized intra and extracellularly. The effect of different pH values, carbon sources and the duration of cultivation time on the gene expression were also studied in liquid and solid media. Results indicated that the produced enzyme had the maximum activity of 41and 38 U/ml for the intra and extracellular, respectively in pHs ranging between 4.5-5.5 at optimum temperature of 30°C after 3 days of yeast culture. Galactose was found to be the most efficient carbon source than the other sources and the maximum activity of target enzyme was o...
Enzyme and Microbial Technology, 2011
Glucose oxidase (GOx) from Penicillium amagasakiense has a higher specific activity than the more commonly studied Aspergillus niger enzyme, and may therefore be preferred in many medical and industrial applications. The enzyme rapidly inactivates on storage at pH 7.0–7.6 at temperatures between 30 and 40°C. Results of fluorimetry and circular dichroism spectroscopy indicate that GOx inactivation under these conditions is
Journal of Chromatography A, 1990
Fast protein liquid chromatography (FPLC) in combination with ion-exchange chromatography on a Mono Q column was used to purify glucose oxidase from Penicillium amagasakiense to homogeneity. Purification was performed with a mixed pH and salt gradient, with 20 mA4 phosphate buffer (pH 8.5) as starting buffer (A) and 50 mM acetate buffer (pH 3.6) with 0.1 M NaCl as elution buffer (B). Elution conditions were optimized to permit the simultaneous purification and separation of the glucose oxidase isoforms. Three peaks, each consisting of l-2 isoforms and exhibiting a homogeneous titration curve profile, were resolved with a very flat linear gradient of 5.0-5.1% B in 40 ml. Three more peaks, each consisting of several isoforms, were eluted at lo%, 30% and 100% B. Optimization of the elution conditions and separation of the glucose oxidase isoforms was only possible because of the rapidity of each purification step and the high resolution provided by FPLC and Mono Q.
Enhancement of glucose oxidase production by Penicillium variabile P16
Enzyme and Microbial Technology, 1999
Effects of the polysaccharides alginate and locust bean gum, and oligosaccharides oligomannuronate OM ( ) ( ) and oligoguluronate OG , on glucose oxidase GOD production by Penicillium variabile P16 were studied. Small increases were obser¨ed when the cultures were supplemented with OG and OM blocks with an a¨erage ( ) y1 degree of polymerization DP of approximately ten. With 200 mg l OM blocks addition at 0 h, the increase ( reached 32.1% compared with the control; howe¨er, regardless of the time of addition, large increases up to ) y 1 approximately 70% in GOD production were obtained with 100 and, particularly, 200 mg l of alginate-de-( ) ri¨ed oligosaccharides OG andr or OM blocks with a DP of approximately se¨en. No significant influence was obser¨ed on mycelial biomass. ᮊ 1999 Else¨ier Science Inc. All rights reser¨ed.
Brazilian Archives of Biology and Technology, 2013
This work aimed to study the production and purification of glucose oxidase by Aspergillus niger and Penicillium notatum using corn steep liquor as the substrate and evaluate its antimicrobial activity for use in pharmaceutical and food industries. The enzyme was purified by ammonium sulfate precipitation (60-85%), DEAE-cellulose ion exchange and Sephadex G-200 size exclusion chromatography. The crude enzyme extracts of A. niger and P.
Glucose oxidase (GO) is an important industrial enzyme typically purified from Penicillium and Aspergillus sp. As GO distribution within the cultures influences process design for maximal product recovery, distribution of GO activity in Penicillium sp. CBS 120262 and Aspergillus niger NRRL-3, during mid-exponential and stationary phases, is compared. On progression from mid-exponential to stationary phase, the percentage GO activity in the cytoplasm decreased 1.6-and 1.3-fold in Penicillium sp. and A. niger respectively. In Penicillium sp., a concomitant 1.8-and 1.9-fold decrease in the percentage GO activity in the cell envelope and slime mucilage respectively, translated into a 2.0-fold increase in the extracellular fluid. In A. niger, decreasing cytoplasmic GO activity was accompanied by 1.3fold increases in the cell envelope and slime mucilage, with a 1.3-fold decrease in the extracellular fluid. Similar trends were observed in specific GO activities. As final GO activity recovered is governed by the purification program, recovery from the extracellular fluid plus cell extract or from the extracellular fluid only were compared through simulating processes of varying complexity. A critical yield for each purification stage was identified above which recovery from the extracellular fluid plus cell extract exceeded that from extracellular fluid alone. These results highlight the influence of microorganism, harvest time and efficiency of downstream process on GO activity delivered. In the systems studied, Penicillium sp. is the organism of choice and should be harvested during stationary phase. The purification process chosen should be informed by both enzyme distribution and individual purification stages yields. Biotechnol. Bioeng. 2008;99: 910-918.
Journal of Chromatography A, 1987
The high resolving power of the preparative and analytical high-performance chromatographic and electrophoretic methods recently developed in this laboratory for the separation of biopolymers has been demonstrated by the purification and characterization of glucose oxidase and catalase from Penicillium chrysogenum. Crude glucose oxidase was purified to homogeneity in one step by high-performance hydrophobic-interaction chromatography (HIC) on a pentylagarose column. Crude catalase was purified by a combination of HIC and high-performance anion-exchange chromatography on 3-diethylamino-2-hydroxypropylagarose. The homogeneity of the enzymes was monitored by high-performance electrophoresis and free zone electrophoresis. The pl values of these two enzymes determined by isoelectric focusing in the high-performance electrophoresis apparatus were 4.2 and 6.5, respectively. Their molecular weights were determined by high-performance molecular sieve chromatography on an agarose column. Glucose oxidase has a molecular weight of 175 000 and probably consists of two identical subunits, as sodium dodecyl sulphate polyacrylamide gel electrophoresis gave a molecular weight of around 72000. The molecular weight of catalase, which is probably composed of non-identical subunits, as indicated by sodium dodecyl sulphate electrophoresis, is around 320 000. Some other characteristics of these two enzymes were also investigated, e.g., electrophoretic mobility, pH stability and optimum pH.