Pectinase Production Research Articles

Production and Purification of Pectinase from Bacillus subtilis 15A-B92 and Its Biotechnological Applications.

Enzymes that degrade pectin are called pectinases. Pectinases of microbial origin are used in juice clarification as the process is cost-effective. This study screened a pectinase-producing bacterium isolated from soil and identified as Bacillus subtilis 15A B-92 based on the 16S rRNA molecular technique. The purified pectinase from the isolate showed 99.6 U/mg specific activity and 11.6-fold purity. The molecular weight of the purified bacterial pectinase was 14.41 ± 1 kD. Optimum pectinase activity was found at pH 4.5 and 50 °C, and the enzyme was 100% stable for 3.5 h in these conditions. No enzymatic inhibition or activation effect was seen with Fe2+, Ca2+, or Mg2+. However, a slight inhibition was seen with Cu2+, Mn2+, and Zn2+. Tween 20 and 80 slightly inhibited the pectinase, whereas iodoacetic acid (IAA), ethylenediaminetetraacetate (EDTA), urea, and sodium dodecyl sulfate (SDS) showed potent inhibition. The bacterial pectinase degraded citrus pectin (100%); however, it was inactive in the presence of galactose. With citrus pectin as the substrate, the Km and Vmax were calculated as 1.72 mg/mL and 1609 U/g, respectively. The high affinity of pectinase for its substrate makes the process cost-effective when utilized in food industries. The obtained pectinase was able to clarify orange and apple juices, justifying its application in the food industry.

Production of pectinases towards the extraction of natural pectin from orange peels using fungal sources

Pectin is a polysaccharide, which is present in middle lamella of cell wall of higher plants. Almost all fruits have a part of pectin in their cell walls. Pectinases are used in the degradation of the plant material and promptly extract juices from different fruits. For this purpose, identified fungi were screened qualitatively and quantitatively for the production of pectinase at 30 ºC for 92 hours of incubation time. In liquid state fermentation, identified strains of Trichoderma asperellum, Aspergillus niger and Thermomyces were treated in which Trichoderma asperellum produced maximum pectinase activities (9.2 U/ml) after 24 hours of incubation by using orange peels as a sole carbon source. Furthermore, pectinases produced from fungal strains were used to produce natural pectin from fruit waste like orange peels. During fermentation process, pectinase produced from fungal strains were inoculated to dried orange peels for 24 hours of incubation time at 30 ºC. The filtered extract was treated with 3 volumes of 80% ethanol for washing to extract pectin. Trichoderma asperellum gave maximum pectin yield [(11.7%)], Aspergillus niger and Thermomyces gave minimum pectin yield [(9.7%, 7.5%)] from orange peels respectively and the amount of pectin yield extracted from orange peels control was [(3.5%)]. Pectinases and pectin has found huge industrial applications in soft drinks, dairy products, pharmaceuticals and food industries, therefore, the present work has focused on the cost effective production of pectinase enzyme and extraction of natural pectin by using potential indigenous fungal strains.

New approach for screening of microorganisms from Arabica coffee processing for their ability to improve Arabica coffee flavor.

The purpose of this research was to isolate microorganisms from coffee fermentation processes and screen them for their potential to improve the flavor of Arabica coffee using a new approach that included pectin degradation ability and growth in mucilage broth. All of the studied microorganisms were isolated from 38 different samples of fresh coffee cherries, coffee mucilage and coffee pulp. A total of 262 microbial isolates were obtained and subjected to screening using pectinase screening agar medium for pectinolytic organisms. The results of the pectinase production test showed that 18 yeast isolates were found to produce pectinase that could degrade the pectin present in solid media. The sugar assimilation profiles and growth of selected strains in mucilage broth were studied. Therefore, 18 isolates from the selected yeasts were subjected to molecular identification by the use of 18S rRNA gene sequencing. The diversity of the yeast isolates was studied, and they were identified as Wickerhamomyces anomalus, Naganishia liquefaciens, Pichia kudriavzevii, Kazachstania naganishii and Kazachstania sp. Moreover, isolates SWU3YWP1-3, SWU3YSK9 and INFCY1-4 were used as a seed culture for Arabica coffee fermentation. The cupping sensory scores of the control (without yeast inoculation) and those inoculated with three isolated yeast strains that were determined by Q-Arabica Graders were 73.75, 84.75, 80.25 and 75.00, respectively. Unique flavors and aromas were detected. This is the first report of screening microorganisms from the Arabica coffee fermentation process by the combination of various properties with success in improving the quality of coffee beverage.

Isolation and characterization of pectinase-producing bacteria (Serratia marcescens) from avocado peel waste for juice clarification

BackgroundBacterial pectinase is an enzyme that could be employed in numerous sectors to break down pectin polysaccharide compounds. The goal of this study is to find pectinase-producing bacteria in avocado peel waste and see if the pectinase enzyme produced can be used to make fruit juice clarification.ResultsThe researchers isolated four different bacterial strains from avocado peel waste samples. The potential two bacterial isolates that were identified as being Serratia marcescens and Lysinibacillus macrolides. Finally, the analysis of pectinase production and its application in fruit juice clarification were performed using one of the bacterial strains of Serratia marcescens. The clear apple, lemon, and mango juices were further processed to assess each juice's properties. The highest antioxidant activity was recorded in lemon juice samples. The lemon juice showed the highest total titratable acidity and total phenol content. Apple juices contained the highest total soluble solids, reducing sugar content, and viscosity and the mango juices have the maximum pH value recorded.ConclusionsThe pectinase isolated from the bacterium Serratia marcescens could clear fruit juices. This pectinase needs to be studied more to make sure it works better in the fruit industry and other businesses.

Response-Surface Statistical Optimization of Submerged Fermentation for Pectinase and Cellulase Production by Mucor circinelloides and M. hiemalis

Cellulase and pectinase are degrading cellulosic and pectic substances that form plant cell walls and, thereby, they have a wide range of applications in the agro-industrial by-products recycling and food industries. In the current research, Mucor circinelloides and M. hiemalis strains were tested for their ability to produce cellulase and pectinase from tangerine peel by submerged fermentation. Experiments on five variables: temperature, pH, incubation period, inoculum size, and substrate concentration, were designed with a Box–Behnken design, as well as response surface methodology (RSM), and analysis of variance was performed. In addition, cellulase and pectinase were partially purified and characterized. At their optimum parameters, M. circinelloides and M. hiemalis afforded high cellulase production (37.20 U/mL and 33.82 U/mL, respectively) and pectinase (38.02 U/mL and 39.76 U/mL, respectively). The partial purification of M. circinelloides and M. hiemalis cellulase produced 1.73- and 2.03-fold purification with 31.12 and 32.02% recovery, respectively; meanwhile, 1.74- and 1.99-fold purification with 31.26 and 31.51% recovery, respectively, were obtained for pectinase. Partially purified cellulase and pectinase from M. circinelloides and M. hiemalis demonstrated the highest activity at neutral pH, and 70 and 50 °C, for cellulase and 50 and 60 °C, for pectinase, respectively. Moreover, 10 mM of K+ increased M. circinelloides enzymatic activity. The production of cellulase and pectinase from M. circinelloides and M. hiemalis utilizing RSM is deemed profitable for the decomposition of agro-industrial wastes.

Inoculation conditions improved the pectinase productivity in Aspergillus niger LB-02-SF solid-state cultivation

In this work, the pectinase production by Aspergillus niger LB-02-SF was evaluated in relation to the inoculation conditions of solid-state cultivations carried out in Becher flasks and in rotating drum bioreactor. From the inoculation by spore suspension in the concentration of 7 × 107 spore/100 g of wet medium (gwm), which is standard inoculum condition, were evaluated: different concentrations of spore suspension (from 7 × 104 to 7 × 108 spore/100 gwm), solid vegetative inoculum using 50 and 100 gwm of fermented solid medium (obtained from 48 h of A. niger cultivation in Becher flasks, in a humidified atmosphere, 30 °C) and liquid vegetative inoculum in the proportions of 5 and 10% (v/w) (obtained from 18 h of A. niger cultivation at 300 rpm, 28 °C). In comparison to the standard condition, with the use of the lowest spore concentration (7 × 104 spore/100 gwm), 100 gwm of solid vegetative inoculum and 10% (v/w) of liquid vegetative inoculum, higher pectinases productivities were observed, around 48, 42 and 85%, respectively. The results obtained indicate that the three inoculation strategies defined are alternatives to increase the enzyme productivity and to facilitate the inoculum preparation stage aimed at applying this process on an industrial scale.

Constructing and expressing acyl-homoserine lactone lactonase (aiiA) gene for enhancing Solanum tuberosum resistance against soft rot disease

Abstract. Suryanti E, Rusamana I, Wahyudi AT, Akhdiya A, Sukamdjaja D. 2022. Constructing and expressing acyl-homoserine lactone lactonase (aiiA) gene for enhancing Solanum tuberosum resistance against soft rot disease. Biodiversitas 23: 1780-1787. Expression of the genes associated with pectinase production in soft rot disease of Dickeya spp. are regulated by quorum sensing mechanism using acyl homoserine lactone(AHL). Degradation of AHL using AHL lactonase can be developed to reduce pectinase production in Dickeyaspp.The aiiA gene encodes AHL lactonase from Bacillus spp. This study aims to construct binary vector of inserted aiiA gene (pCAB10) from Bacillus cereus B10 and express the aiiA gene in potato Medians cultivar genome. The aiiAgene was cloned into binary vector and transformed into Agrobacterium tumefaciens LB4404. The Agrobacterium-mediated genetic transformation was conducted with co-cultivation technique.The transgenic plant was confirmed through PCR andselected through kanamycin and soft rot disease resistance to Dickeya dadantii A3. The result showed that aiiA gene was successfully integrated top CAB10 and introduced to A. tumefaciens LB4404. The percentage of pCAB10 transformation was 57.6%, while there generation efficiency was 11.9% with 50 mg/L acetosyringone addition. Four transgenic potato Medians cultivar exhibited significantly higher resistance against soft rot disease than non-transgenic plants. Mahl1 clone had the highest resistance to D. dadantii with soft rot disease incidence as 7.1% and percentage of soft rot disease suppression as 92.85%.

Multi-Objective Statistical Optimization of Pectinolytic Enzymes Production by an Aspergillus sp. on Dehydrated Coffee Residues in Solid-State Fermentation

Pectinolytic enzymes are a group of enzymes widely used in the food industry. They can be obtained through a wide range of by-products and agricultural and agro-industrial waste by the action of fungi, such as Aspergillus spp., by solid-state fermentation (SSF). A wild strain of an Aspergillus sp. isolated in the Universidad Técnica del Norte (UTN) was used to obtain pectinolytic enzymes from dehydrated coffee waste (pulp and husk) derived from coffee cherries cultivated in the Ecuadorian Andean regions. It was possible to find a condition in which the production of pectinases (expressed as Enzymatic Activity (EA)) and the concentration of spores (S) were simultaneously maximized, using the response surface methodology, in a 3-level factorial design, by SSF in simple tray-type bioreactors. After the analysis and optimization of quadratic models, three confirmatory experiments were performed in the unique optimal condition recommended (35 °C and 79% relative humidity), obtaining 29.9 IU/g and 2.64 × 106 #Sp./g for EA and S, respectively; these values coincided with those predicted by the quadratic models, demonstrating their validity. The values obtained in this study are similar to those previously obtained by other authors.

SCREENING OF PECTINASE PRODUCING BACTERIA FROM SOIL AND PRODUCTION, PARTIAL PURIFICATION OF ENZYME

Bacterial pectinase enzyme is very much important at industrial level for different processes This present study focus on bacterial pectinase. Pectinase producing bacterial spp.is screen out from soil. About seven cultures of bacterial spp. were isolated from farmland soil sample using pectin agar plates. The selected isolates showed the zone of hydrolysis to confirmed the production of pectinase from the isolates. Among them the culture S4 showed prominent zone of hydrolysis surrounding the colony. Pectinase activity of each isolate were checked at an interval of 24 hrsusing DNSA method to access the enzyme efficiency produced by respective isolates. Keeping the ideal growth conditions the highest pectinase activity obtained among isolates was 11.43 U/ ml, by culture S4.Optimum condition for pectinase production was found at 72hrs. incubation period, 37˚C temperature and alkaline pH of 10 could be the optimum cultural conditions for bacterial strain S4. Bacterial strain S4 gave 40.85 U/ml partially purified enzyme.

SCREENING FOR PECTINASE PRODUCTION IN SOLID STATE FERMENTATION (SSF) OF BANANA PEELS BY BACILLUS SPP

Solid state fermentation of banana peels by Bacillus spp for pectinase production was investigated using standard methods. Optimum conditions that affect the growth and pectinase production by the microorganism such as incubation temperature ( 35OC), inoculum size ( 3.00mL), incubation period (24h), pH (7), substrate concentration (30g) and additional nitrogen source (0,30%) were determined and employed in the fermentation . Results of enzyme production measured as enzyme activity at these optimum growth conditions were 7.00, 4.00, 3.90, 4.10,3,50 and 3.50 U/mL/min. respectively which were the highest value in each case. Further increase or decrease in growth conditions above the optimum in all cases did not cause any increase in pectinase activity. The finding suggests that the various optimum growth conditions for Bacillus spp in solid state fermentation of banana peels can be of economic value in the production of pectinase, a very useful industrial enzyme. The procedure can also be employed in getting rid of surplus wastes that would otherwise accumulate from large scale cultivation of banana as currently practiced all over the world including Nigeria.

Concurrent production of cellulase, xylanase, pectinase and immobilization by combined Cross-linked enzyme aggregate strategy- advancing tri-enzyme biocatalysis

In this study Bacillus sp. isolated from snail droppings displayed ability to coproduce cellulase, xylanase and pectinase by solid state fermentation on citrus peel. The enzymes were immobilized by cross-linking enzyme aggregate (CLEA). Parameters like cross-linking concentration/cross-linking time, pH, thermal stability and reusability were evaluated. Bacillus sp. expressed cellulase (2.563 ± 0.082 U/gds), xylanase (2.253 ± 0.101 U/gds) and pectinase (2.910 ± 0.097 U/gds) activity. Moisture content (75%), pH 5, particle size 348 nm, One (1 mL) mililiter inoculum/nitrogen were best for enzymes production. Optimum aggregation of enzyme complex with glutaraldehyde concentration (50 mM) and cross-linking time 3 h was achieved using three-phase portioning (TPP). Combined-CLEAs expressed thermal stability. Cellulase, xylanase and pectinase, retained 90% of initial activity at 1 h, over 65% after 3 h incubation at 50 °C; optimum activity was pH 6. After 4 cycles combined-CLEAs retained over 80% activity.

Pectinase from Microorganisms and Its Industrial Applications.

The utilization of microbial pectinase in different industries has been increased in its world demand. The major sources of pectinase are microorganisms mainly bacteria, fungi and yeast. The utilization of low-cost agro-industrial wastes as substrates has been preferable in pectinase production. Pectinase production faced various parameters optimization constraints such as temperature, pH and production times which are the main factors in pectinase production. The pectinase enzyme is getting attention due to its several advantages; hence, it needs to be explored further to take its maximum advantage in different industries. This review discusses the pectin substance structure, substrate for pectinase production, factors influencing pectinase production, the industrial application of microbial pectinase and also discusses challenges and future opportunities of applying microbial pectinase in industry.

Optimisation of Xylanase–Pectinase Cocktail Production with Bacillus amyloliquefaciens ADI2 Using a Low-Cost Substrate via Statistical Strategy

An effective statistical tool for increasing and boosting the production of xylanase and pectinase by Bacillus amyloliquefaciens ADI2 during submerged fermentation (SmF) appears to be the response of surface methodology (RSM) using the central composite design (CCD). Optimum production was achieved under fermentation conditions of a temperature of 28 °C, pH of 8.38, inoculum size of 4% (w/v) and agitation speed of 94 rpm for 48 h. The experimental responses demonstrated a near agreement with the expected responses under optimum conditions of independent variables, suggesting the model’s validity. The optimised CCD model had a 1.34-fold, 159 ± 6 U/mL greater xylanase and 5.96-fold, 205 ± 9 U/mL greater pectinase production than the one factor at a time (OFAT) approach. The production of concurrent enzymes of xylanase–pectinase resulted in a ratio of 1:1.3.

Exploring the phenotypic diversity of oenological traits in Kluyveromyces marxianus strains.

The use of non-Saccharomyces yeasts in the winemaking process may have several positive outcomes. Kluyveromyces marxianus has recently been revealed as a promising species for this industry. While the majority of studies mention the use of K. marxianus in various industries including food production (e.g. dairy and cocoa), recent studies have also shown that its aroma and pectinase production make it a suitable yeast for the wine industry. Nevertheless, only particular strain, IWBT Y885, was investigated. In this study, five different K. marxianus strains as well as one protoplast fusant (BF2020) were compared to strain Y885. These comparisons focused on various oenological traits such as fermentation performance, fermentation metabolites, hydrogen sulfide, and pectinase production. Throughout the study, variations were found between the K. marxianus strains investigated. Indeed, although common traits such as high pectinase activity appeared conserved among K. marxianus strains, a fairly large phenotypic diversity was also evident. Using cluster analysis, strain groupings emerged with strains L01, L05, Y885, and BF2020 grouping together. Similarly, strains L02 and L04 grouped together while strain L03 appearing to show the most variation between the strains investigated. Variation between strains was observed regardless of the original source of isolation.

Heterogenicity within the LPS Structure in Relation to the Chosen Genomic and Physiological Features of the Plant Pathogen Pectobacterium parmentieri.

Pectobacterium parmentieri is a pectinolytic plant pathogenic bacterium causing high economic losses of cultivated plants. The highly devastating potential of this phytopathogen results from the efficient production of plant cell wall-degrading enzymes, i.e., pectinases, cellulases and proteases, in addition to the impact of accessory virulence factors such as motility, siderophores, biofilm and lipopolysaccharide (LPS). LPS belongs to pathogen-associated molecular patterns (PAMPs) and plays an important role in plant colonization and interaction with the defense systems of the host. Therefore, we decided to investigate the heterogeneity of O-polysaccharides (OPS) of LPS of different strains of P. parmentieri, in search of an association between the selected genomic and phenotypic features of the strains that share an identical structure of the OPS molecule. In the current study, OPS were isolated from the LPS of two P. parmentieri strains obtained either in Finland in the 1980s (SCC3193) or in Poland in 2013 (IFB5432). The purified polysaccharides were analyzed by utilizing 1D and 2D NMR spectroscopy (1H, DQF-COSY, TOCSY, ROESY, HSQC, HSQC-TOCSY and HMBC) in addition to chemical methods. Sugar and methylation analyses of native polysaccharides, absolute configuration assignment of constituent monosaccharides and NMR spectroscopy data revealed that these two P. parmentieri strains isolated in different countries possess the same structure of OPS with a very rare residue of 5,7-diamino-3,5,7,9-tetradeoxy-l-glycero-l-manno-non-2-ulosonic acid (pseudaminic acid) substituted in the position C-8: →3)-β-d-Galf-(1→3)-α-d-Galp-(1→8)-β-Pse4Ac5Ac7Ac-(2→6)-α-d-Glcp-(1→6)-β-d-Glcp-(1→. The previous study indicated that three other P. parmentieri strains, namely IFB5427, IFB5408 and IFB5443, exhibit a different OPS molecule than SCC3193 and IFB5432. The conducted biodiversity-oriented assays revealed that the P. parmentieri IFB5427 and IFB5408 strains possessing the same OPS structure yielded the highest genome-wide similarity, according to average nucleotide identity analyses, in addition to the greatest ability to macerate chicory tissue among the studied P. parmentieri strains. The current research demonstrated a novel OPS structure, characteristic of at least two P. parmentieri strains (SCC3193 and IFB5432), and discussed the observed heterogenicity in the OPS of P. parmentieri in a broad genomic and phenotype-related context.

Creating wealth from waste: An approach for converting organic waste in to value-added products using microbial consortia

In the current scenario, great emphasis is being made on the recovery, recycling, and reconditioning of organic solid waste. With a focus on the bioconversion of organic waste into value-added products, municipal vegetable waste was fractionated into liquid and solid part. The solid fraction was subjected to biodegradation in the presence of different microbial consortia and different enzymes were extracted. The remaining solid stuff was converted into animal feed, followed by the evaluation of its microbiological and chemical indices. Liquid fraction separated from waste was used as a crude medium to grow different bio-fertilizer strains. Among the three consortia, consortium C (Aspergillus terreus and Myrothecium verrucaria) showed the maximum production of amylase, cellulase, xylanase and pectinase after 4 days. Results of the microbiological and chemical parameters of the animal feed prepared from the fermented vegetable waste showed that the various parameters were found to be within the permissible limit. The average concentrations of heavy metals in the compost was found to be within the permissible limit of FCO standard, therefore, it can also be used for agricultural application. For using liquid extract of the vegetable waste, maximum CFU count 1.9 × 106, 1.1 × 106, and 8.7 × 105 was achieved in BAB 1980, BAB 662, and BAB 267 strain, respectively after 30 h of incubation. Thus, the present study demonstrated the bioconversion of vegetable waste into crude enzymes, animal feed, compost, and liquid biofertilizer.

Isolation, screening, and molecular identification of pectinase producers from fruits, vegetables, and soil samples

Background and objective In this work, isolation, identification (morphological and chemical), and molecular characterization were done of local isolates of some pectinase-producing microorganisms such as bacteria, actinomycetes, fungi, and yeast.Materials and methods A total of 22 local bacterial isolates were obtained from various sources and were assayed for pectinolytic activity after optimization of conditions for pectinase production. Isolate no. 19 showed the highest pectinase-specific activity (6.73 U/ml) on glucose-supplemented medium, whereas isolate no. 5 gave the lowest pectinase productivity (3.21 U/ml). The identification of isolate no. 19 revealed that it belonged to the genus Bacillus based on morphological and biochemical characteristics. Based on molecular identification (16 S rRNA technique), isolate no. 19 was named Bacillus sp. strain NRBANKI-4 (with 99% similarity), with Gene Bank accession number OM540351.Results and conclusion A total of 14 local actinomycete isolates were obtained from soil samples. Isolate no. 13 showed the highest pectinase-specific activity (6.48 U/ml), whereas sample no. 10 gave the lowest pectinase-specific activity (3.07 U/ml). Based on molecular identification (16 S rRNA technique), isolate no. 13 was named Streptomyces sp. KP 12 (90.63% similarity), with Gene Bank accession number OM403596. A total of 10 fungal isolates were obtained from crop waste soil. Isolate no. 2 gave the highest pectinase productivity (21.20 U/ml). Based on molecular identification (internal transcribed spacer-PCR technique), isolate no. 2 was named Aspergillus niger F8121 (99.47% similarity), with Gene Bank accession number OM392061. Following the same trend, 10 yeast isolates were isolated from crop waste soil. The isolate that gave the highest pectinase productivity was no. 7, which gave 22.03 U/ml. The isolate that gave the lowest was no. 9 (20.74 U/ml). Isolate no. 7 was named Pichia barkeri Y1 (90.91% similarity), with Gene Bank accession number OM392066.

Insights of Pectinase from Fungi

The plant’s cell walls contain pectin, a gelatinous heteropolysaccharide that is integrated into the cell and other parts of the cell wall that allow cells to co-exist and are essential for plant growth. Enzymes called pectinase serves as an important agent in the breakdown of the components of pectin substances. Pectinase is used to degrade the pectin present in vegetables and thus helps to reinforce fruit juice extraction from fruits such as apples, tomatoes, and oranges. The current study aims to isolate, screen, and optimize the pectolytic enzyme from fungi of environmental samples. Twenty of the 25 isolates showed a zone of hydrolysis, indicating an organism’s ability to produce pectinase enzyme. By performing screening of pectinase, 80% of positive isolates were obtained, and these isolates were optimized to achieve high Pectinase production by carrying out various parameters such as pH, temperature, and carbon source, and it was found that pH 5, 30 oC and 2% of carbon source were found to be optimum for the production of pectolytic enzyme. The molecular weight of the isolated enzyme was estimated by sodium dodecyl sulphate polyacrylamide gel electrophoresis, and was determined to be approximately 66 kDa.

Development and Validation of fermentation conditions for Pectinase production in submerged fermentation conditions

Fermentation technology plays a very important role in the field of Biotechnology. Production of various enzymes, antibiotics, single cell proteins, various food products, etc is carried out in a vessel called as the bioreactor which is the seed of fermentation technology. The project includes study of Pectinase production using submerged fermentation and various parameters like different substrate, different strains and fermentation time to obtain the maximum yield of Pectinase. To achieve this various research is being conducted. Bacillus subtilis species was being used in this work for the production of Pectinase.

EXTRACELLULAR ALKALINE PECTINASES PRODUCTION: A REVIEW

Pectinases are most significant enzymes of commercial sector. Among all the commercially produced industrial enzymes, pectinases occupy 25% share in the overall enzyme market. Pectinases have wide applications in various processes such as fruit juice clarification, wastewater remediation and textile and paper industry etc. Pectinase is most suitable environment friendly tool of nature for sustainable development. Although, pectinases alike many other commercial enzymes are facing the imperative of low harvest and yield. Hence, pectinases have been the objective of research to accomplish the commercial scale articulation levels. To find unique physicochemical properties of pectinases often microbial pectinases are utilized among all the natural sources due to ease in production. Present study primarily centers on the different types of recent techniques for the alkaline pectinases production and optimization strategies advancement for high production yield. For achieving higher production of the extracellular alkaline pectinases by utilizing basic fermentation media and simple molecular devices along with genetic engineering approaches are included in the present study. For the improved alkaline pectinase production advanced genetic engineering techniques such as gene cloning, tools of r-DNA technology and cell immobilization technique are discussed.