Production of cold-active pectinases by three novel Cladosporium species isolated from Egypt and application of the most active enzyme

Abstract

Cladosporium parasphaerospermum, Cladosporium chlamydosporigenum, and Cladosporium compactisporum have all been discovered and characterized as new Cladosporium species. The three new species seemed to generate cold-active pectinases with high activity at pH 6.0 and 10 °C, pH 6.0 and 15 °C, and pH 5.0 and 15 °C, respectively, with the most active being C. parasphaerospermum pectinase. In submerged fermentation (SmF), C. parasphaerospermum produced the most cold-active pectinase with the highest activity and specific activity (28.84 U/mL and 3797 U/mg) after 8 days. C. parasphaerospermum cold-active pectinase was isolated using DEAE-Cellulose anion exchange resin and a Sephadex G 100 gel filtration column. The enzyme was purified 214.4-fold and 406.4-fold greater than the fermentation medium using DEAE-cellulose and Sephadex G 100, respectively. At pH 7.0 and 10 °C, pure pectinase had the highest activity (6684 U/mg), with Km and Vmax determined to be 26.625 mg/mL and 312.5 U/min, respectively. At 5 mM/mL, EDTA, MgCl2, and SDS inhibited the activity of pure pectinase by 99.21, 96.03, and 94.45%, respectively. The addition of 10 U/mL pure pectinase enhanced the yield of apple, orange, apricot, and peach juice by 17, 20, 13, and 24%, respectively, and improved the clarity and colour of orange juice by 194 and 339%, respectively. We can now add cold-active pectinase production to the long list of Cladosporium species that have been identified. We also report three new species that can be used in biotechnological solutions as active microbial pectinase producers. Although further research is needed, these distinct species might be used to decompose difficult and resistant pertinacious wastes as well as clear fruit juices.

Production of cocktail enzymes and partial purified pectinase utilizing lignocellulosic wastes by specific Didymella species

“…AUMC 15636) was used to create pectinase enzyme in SmF. Pectinase has been used for a long time to degrade pectin and speed up various processing procedures, such as juice extraction, clarification, and liquefaction [16,56]. Pectinases are one of the most used enzymes in food, accounting for 40% of all dietary enzymes [57,58].…”

“…Total protein from 1.0 L fermentation medium was isolated from the solution at 4 ºC using triple volume cold ethanol (96.0 %), and dried by freeze dryer (VirTis, model # 6 KBTES-55, NY, USA). A 0.9 g enzyme powder was dissolved in 5 mL 100 mM citrate buffer (pH 5.0) and dialyzed according to Moharram et al [16], and utilized as partly pure fungal pectinase in characterization tests.…”

“…These enzymes break down pectin, a complex polymer hold the cellulose network together. Pectinases make almost tenth of all commercial enzymes produced worldwide, and their market is expanding daily [13][14][15][16]. In order to produce cocktail enzymes by three Didymella species in SSF and partially purify pectinase enzyme in SmF, the current research aims to employ some remaining lignocellulosic wastes as an alternative carbon source.…”

Cold‐active microbial enzymes and their biotechnological applications

“…These enzymes are distinguished by their optimum catalytic activity at temperatures between those considered low and adequate; nevertheless, they are also heat‐sensitive and rapidly inactivated over a certain temperature (Hamid et al., 2022 ). Due to their high efficiency and environmentally friendly nature, enzymes are in great demand in many industrial applications, including food and beverage production (Grzonka et al., 2007 ; Moharram et al., 2022 ; Schäfer et al., 2006 ; Singh et al., 2016 ). Moreover, these enzymes are an essential component of a well‐established global sector that is projected to increase in value over the next several years to US$6.3 billion (Dewan, 2017 ; Moharram et al., 2022 ; Singh et al., 2016 ).…”

“…Due to their high efficiency and environmentally friendly nature, enzymes are in great demand in many industrial applications, including food and beverage production (Grzonka et al., 2007 ; Moharram et al., 2022 ; Schäfer et al., 2006 ; Singh et al., 2016 ). Moreover, these enzymes are an essential component of a well‐established global sector that is projected to increase in value over the next several years to US$6.3 billion (Dewan, 2017 ; Moharram et al., 2022 ; Singh et al., 2016 ). These days, it's common practice to use cold‐active enzymes to bring down the temperature of industrial processes.…”