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. 2022 Jun 29;27(13):4195.
doi: 10.3390/molecules27134195.

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

Yahya S Alqahtani  1 , Sunil S More  2 , Keerthana R  2 , Ibrahim Ahmed Shaikh  3 , Anusha K J  2 , Veena S More  4 , Francois N Niyonzima  5 , Uday M Muddapur  6 , Aejaz A Khan  7
Affiliations

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

Yahya S Alqahtani et al. Molecules. .

Abstract

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.

Keywords: Bacillus subtilis 15A-B92; citrus pectin; juices clarification; pectinase; purification; screening.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Bacterial isolate showing zone of hydrolysis after iodine solution application.
Figure 2
Figure 2
Phylogenetic tree showing the position of isolated bacterium (AK).
Figure 3
Figure 3
SDS PAGE pattern of purified pectinase purified from Bacillus subtilis 15A-B92.
Figure 4
Figure 4
Deconvoluted mass spectrum of bacterial pectinase from Bacillus subtilis 15A-B92.
Figure 5
Figure 5
Juice clarification. (A): unclarified and clarified apple juice. (B): unclarified and clarified orange juice.
See this image and copyright information in PMC

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