Phylogenetic Analysis of Polygalacturonase-Producing Bacillus and Pseudomonas Isolated From Plant Waste Material

Abstract

Background:Keeping in mind the commercial application of polygalacturonase (PG) in juice and beverages industry, bacterial strains were isolated from rotten fruits and vegetables to screen for competent producers of PG.Objectives:In this study, the plate method was used for preliminary screening of polygalacturonase-producing bacteria, while the Dinitrosalicylic Acid (DNS) method was used for quantifications of PG.Materials and Methods:Biochemically-identified polygalacturonase-producing Bacillus and Pseudomonas species were further characterized by molecular markers. The genetic diversity among these selected strains was analyzed by investigating microsatellite distribution in their genome. Out of 110 strains, 17 competent strains of Bacillus and eight strains of Pseudomonas were selected, identified and confirmed biochemically. Selected strains were characterized by 16S rRNA sequencing and data was submitted to the national center for biotechnology information (NCBI) website for accession numbers.Results:Among the Bacillus, Bacillus vallismortis (JQ990307) isolated from mango was the most competent producer of PG; producing up to 4.4 U/µL. Amongst Pseudomonas, Pseudomonas aeruginosa (JQ990314) isolated from oranges was the most competent PG producer equivalent to B. vallismortis (JQ990307). To determine genetic diversity of different strains of Pseudomonas and Bacillus varying in PG production, fingerprinting was done on the basis of Simple Sequence Repeats (SSR) or microsatellites. The data was analyzed and a phylogenetic tree was constructed using the Minitab 3 software for comparison of bacterial isolates producing different concentrations of PG. Fingerprinting showed that presence or absence of certain microsatellites correlated with the ability of PG production.Conclusions:Bacteria from biological waste were competent producers of PG and must be used on an industrial scale to cope with the demand of PG in the food industry.

Evaluation of process involved in the production of aromatic compounds in Gram‐negative bacteria isolated from vanilla ( Vanilla planifolia ex. Andrews) beans

“…By this analysis, the activity from Pseudomonas sp. IFVB was lower in comparison to other Pseudomonas species (4Á4 AE 0Á1 U µl À1 ) isolated from vegetable material (Sohail and Latif 2016). While, activities from Enterobacter sp.…”

LC/MS detection of oligogalacturonic acids obtained from tragacanth degradation by pectinase producing bacteria

“…Bacillus vallismortis is a Gram-positive endospore forming bacterium very close to B. subtilis [29]. The ability of utilizing several aromatic pollutants such as naphthalene, pyrene, phenol and cyclohexane, fibrinolytic and pectinolytic activities have been reported in B. vallismortis [30,31]. The exopolygalacturonate lyase gene was also reported in the complete genome sequence of B. vallismortis TD3 (NZ_NXEM01000029.1) in NCBI.…”

Review on bacterial production of alkaline pectinase with special emphasis on Bacillus species

“…Chessen et al, 1980 reviewed that bacteria like Bacillus, Pseudomonas and Micrococcus isolated from retting fl ax, jute, sisal and coir, and Erwinia from coffee fruits have shown to possess the ability to degrade pectin by producing pectinolytic enzymes (Chesson, 1980). Other bacterial genera reported to have pectinolytic properties include species of Pseudomonas (Sohail & Latif, 2016), -Tapias et al, 2015) Lactobacillus (Karam & Belarbi, 1995) etc.…”

“…It has been shown that bacteria produce pectinase that withstands high pH and temperature (Hoondal et al, 2002;Andrade et al, 2011). Also, it is easy to harvest pectinase than fungus as it is an extracellular product in bacterial culture (Sohail & Latif, 2016). Bacterial strains producing commercial enzymes are always preferred over fungal strains because of ease of fermentation process (for production) and implementation of strain improvement techniques or any modern technique to increase the production yield (Prathyusha & Suneetha, 2011).…”

“…makeup about 50 % of the total enzyme market (Schallmey et al, 2004). Another major feature that makes these groups predominant is that most of them are environmental friendly, don't have fastidious nutritional requirements and are easy to grow and handle (Sohail & Latif, 2016). Bacillus species such as B. subtilis and B. licheniformis are on the Food and Drug…”