Gene cloning, functional expression and characterisation of a novel type I pullulanase from Paenibacillus barengoltzii and its application in resistant starch production

Protein Expression and Purification
Jingjing LiuQiaojuan Yan

Abstract

A novel pullulanase gene (PbPulA) from Paenibacillus barengoltzii was cloned. PbPulA has an open reading frame of 2028 bp encoding 675 amino acids. It was heterologously expressed in Escherichia coli as an intracellular soluble protein. The recombinant pullulanase (PbPulA) was purified to homogeneity with a molecular mass of about 75 kDa on SDS-PAGE. PbPulA was optimally active at pH 5.5 and 50 °C. It was stable within pH 5.5-10.5. The enzyme exhibited strict substrate specificity towards pullulan, but showed relatively low activity towards amylopectin and no activity towards other tested polysaccharides. The Km and Vmax values of the enzyme on pullulan were 2.94 mg/mL and 280.5 μmol/min/mg, respectively. The addition of PbPulA in gelatinized rice and maize starches significantly increased the resistant starch type 3 (RS3) yields. Final yields from rice and maize starches were 10.82 g/100 g and 11.41 g/100 g, respectively. These properties make PbPulA useful in starch industries.

References

Feb 19, 2000·Journal of Chromatography. B, Biomedical Sciences and Applications·G Kriegshäuser, W Liebl
Jul 9, 2004·Critical Reviews in Microbiology·Monika Domań-Pytka, Jacek Bardowski
Apr 4, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Dominggus MalleBunzo Mikami
Sep 20, 2012·Enzyme Research·Siew Ling HiiArbakariya Bin Ariff
Dec 10, 2013·Protein Expression and Purification·Huawei WuGuangxu Wu
Jul 30, 2014·Extremophiles : Life Under Extreme Conditions·Farah QouraGarabed Antranikian

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Citations

May 2, 2020·Acta Crystallographica. Section D, Structural Biology·Ping HuangZhengqiang Jiang
Oct 10, 2019·Anais Da Academia Brasileira De Ciências·Soledad Caminata LandrielHernán Costa
Jun 21, 2021·Biotechnology Advances·Wei XiaJing Wu

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