Pectinase, a group of pectin degrading enzymes, is one of the most influential industrial enzymes, helpful in producing a wide variety of products with good qualities. These enzymes are biocatalysts and are highly specific, non-toxic, sustainable, and eco-friendly. Consequently, both pectin and pectinase are crucially essential biomolecules with extensive applicatory perception in the biotechnological sector. The market demand and application of pectinases in new sectors are continuously increasing. However, due to the high cost of the substrate used for the growth of microbes, the production of pectinase using microorganisms is limited. Therefore, low-cost or no-cost substrates, such as various agricultural biomasses, are emphasized in producing pectinases. The importance and implications of pectinases are rising in diverse areas, including bioethanol production, extraction of DNA, and protoplast isolation from a plant. Therefore, this review briefly describes the structure of pectin, types and source of pectinases, substrates and strategies used for pectinases production, and emphasizes diverse potential applications of pectinases. The review also has included a list of pectinases producing microbes and alternative substrates for commercial production of pectinase applicable in pectinase-based industrial technology. Key points
 • Pectinase applications are continuously expanding. • Organic wastes can be used as low-cost sources of pectin. • Utilization of wastes helps to reduce pollution.

New insights in pectinase production development and industrial applications.

Formulation of the agro-waste mixture for multi-enzyme (pectinase, xylanase, and cellulase) production by mixture design method exploiting Streptomyces sp.

The present study identified the pectinase-producing bacterium isolated from the contaminated broth as Bacillus sp. on 16S rDNA sequence analysis. The bacterium illustrated water-like droplets on the colony grown on the Sabouraud dextrose agar plate. It also exhibited multi-enzymes activities, such as pectinase, polygalacturonase, xylanase, and cellulase by using various agro-wastes as low-cost substrates. The orange peel was observed to be the best substrate among the agro-wastes used for maximum multi-enzymes (pectinase, polygalacturonase, xylanase, and cellulase). However, the bacterium demonstrated its capability to produce different enzymes according to the different substrates/agro-wastes used. The Plackett–Burman design was used to determine the essential influencing factors, while the Box Behnken design response surface methodology was for optimizing cultural conditions. At their optimal conditions (40°C incubation temperature, 24 h of incubation period, 1% w/v orange peel, and 2% v/v inoculum volume), the bacterium exhibited the maximum pectinase (9.49 ± 1.25 U/ml) and xylanase (16.27 ± 0.52 U/ml) activities. Furthermore, the study explored the ability of the bacterium to produce bacterial lipids and observed about 25% bacterial lipid content on a dry weight basis. Therefore, the bacterium is a good candidate for producing important multi-enzymes and subsequent agro-waste degradation controlling the environment, and facilitating waste management. Also, the bacterium can be a potential feedstock in producing renewable biofuel.

/pdf/optimization-of-multiple-enzymes-production-by-fermentation-2wb11rcc.pdf

Optimization of multiple enzymes production by fermentation using lipid-producing Bacillus sp.

Biomass-Degrading Enzyme(s) Production and Biomass Degradation by a Novel Streptomyces thermocarboxydus

Enzyme mediated degradation of lignocellulosic biomass is an important step in waste-biorefineries. Multienzyme preparations can effectively degrade complex materials and, hence, can be applied in biorefineries. Here, an agro-industrial waste, sugarcane bagasse, was used to produce a bacterial multienzyme. The bacterial strains including B. thuringiensis B45, B. velezensis BF3 and B. amyloliquefaciens B987 exhibited their growth at temperatures from 30–50 °C in the presence of 2% salt. The isolates B45, BF3 and B987 were able to produce endoglucanase, xylanase and pectinase, respectively. Therefore, it was aimed to obtain a multienzyme preparation by cultivating the bacterial consortium under a solid-state fermentation of untreated and chemically treated sugarcane bagasse. The results showed that the titres of cellulase and xylanase were generally higher when the strain B45 cultivated at the start of the fermentation. Interestingly, the degradation of cellulose and hemicellulose present in sugarcane bagasse by the strains B45 and BF3 rendered the mere pectin component available to the pectinolytic strain B987. The degradation of SB by the consortium was confirmed by gravimetric analysis and scanning electron microscopy. The study showed that the bacterial strains can be cultivated under solid-state fermentation to obtain industrially important enzymes.

/pdf/preparation-of-a-pectinase-enriched-multienzyme-under-solid-hi1rjiyo.pdf

Preparation of a Pectinase-Enriched Multienzyme under Solid State Fermentation of Sugarcane Bagasse

Pectinases are a group of enzymes with broad application, including in plant fiber processing, pectic wastewater treatment, paper pulping, fruit juice extraction, and clarification. With an increasing industrial demand for these enzymes, it is useful to isolate organisms that produce large amounts of pectinase and possess wide ranges of stability factors like temperature and pH. In this study, 17 out of 29 bacteria (58.62%) from forest soil samples were pectinolytic. However, only four bacteria (S-5, S-10, S-14, and S-17) showed high pectin hydrolysis zones (ranging from 0.2 cm to 1.7 cm). These four bacteria were identified based on colony morphology, microscopic characterization, biochemical characteristics, and 16S rDNA sequencing. They were designated as Streptomyces sp. (S-5, S-14), Cellulomonas sp. (S-10), and Bacillus sp. (S-17). Interestingly, bacteria showed cellulase and xylanase activity in addition to pectinase. The quantitative assay for pectinase activity of the four isolates provided proof that they are pectinase producers and can be considered potential candidates for industrial uses. The crude enzyme extracts of these bacteria are applicable in oil and juice extraction from sesame seeds and apples, respectively.

Screening and Molecular Identification of Novel Pectinolytic Bacteria from Forest Soil

Amylases account for about 30% of the global market of industrial enzymes, and the current amylases cannot fully meet industrial needs. This study aimed to identify a high α-amylase producing bacterium WangLB, to clone its α-amylase coding gene, and to characterize the α-amylase. Results showed that WangLB belonged to Bacillus velezensis whose α-amylase gene was 1980 bp coding 659 amino acids designated as BvAmylase. BvAmylase was a hydrophilic stable protein with a signal peptide and a theoretical pI of 5.49. The relative molecular weight of BvAmylase was 72.35 kDa, and was verified by SDS-PAGE. Its modeled structure displayed that it was a monomer composed of three domains. Its optimum temperature and pH were 70 °C and pH 6.0, respectively. It also showed high activity in a wide range of temperatures (40–75 °C) and a relatively narrow pH (5.0–7.0). It was a Ca2+-independent enzyme, whose α-amylase activity was increased by Co2+, Tween 20, and Triton X-100, and severely decreased by SDS. The Km and the Vmax of BvAmylase were 3.43 ± 0.53 and 434.19 ± 28.57 U/mg. In conclusion, the α-amylase producing bacterium WangLB was identified, and one of its α-amylases was characterized, which will be a candidate enzyme for industrial applications.

Xiaodong Zhang

Papers

Screening and Molecular Identification of Novel Pectinolytic Bacteria from Forest Soil

Bacillus velezensis Identification and Recombinant Expression, Purification, and Characterization of Its Alpha-Amylase