Purification and biochemical characterization of extracellular glucoamylase from Paenibacillus amylolyticus strain

Abstract

In the present study, glucoamylase produced from a soil bacterium Paenibacillus amylolyticus NEO03 was cultured under submerged fermentation conditions. The extracellular enzyme was purified by starch adsorption chromatography and further by gel filtration, with 2.73‐fold and recovery of 40.02%. The protein exhibited molecular mass of ∼66,000 Da as estimated by SDS–PAGE and depicted to be a monomer. The enzyme demonstrated optimum activity at pH range 6.0–7.0 and temperature range 30–40 °C. Glucoamylase was mostly activated by Mn2+ metal ions and depicted no dependency on Ca2+ ions. The enzyme preferentially hydrolyzed all the starch substrates. High substrate specificity was demonstrated towards soluble starch and kinetic values Km and Vmax were 2.84 mg/ml and 239.2 U/ml, respectively. The products of hydrolysis of soluble starch were detected by thin layer chromatography which showed only D‐glucose, indicating a true glucoamylase. The secreted glucoamylase from P. amylolyticus strain possesses properties suitable for saccharification processes such as biofuel production.

Detergent-stable amylase production by Paenibacillus lactis strain OPSA3 isolated from soil; optimization by response surface methodology

“…Paenibacillus contains some of the best-known carbohydrate-active enzyme (CAZyme)-producing bacteria, like P. polymyxa, P. amylolyticus and Paenibacillus thiaminolyticus [ [70] , [71] , [72] , [73] ]. Despite previous reports of amylase production by Paenibacillus species, the current one, to the best of our knowledge, is the first to associate a strain of P. lactis with some extracellular amylase production.…”

Improving Thermostability of Chimeric Enzymes Generated by Domain Shuffling Between Two Different Original Glucoamylases

“…So, searching for a new source of glucoamylase with potentially applicable properties encompassing elevated temperature, extreme pH, high salinity, organic solvents, surfactants, and specificities (substrate and product) is still of considerable importance ( Schmidt et al, 2019 ). Although some novel glucoamylases have been found and characterized for industrial applications ( Guo et al, 2019 ; Karim et al, 2019 ; Lincoln et al, 2019 ; Zhang et al, 2019 ; Wang et al, 2020 ; Lago et al, 2021 ; Wayllace et al, 2021 ), they did not achieve industrially desirable traits.…”

Rational reformation of Corynebacterium glutamicum for producing L-lysine by one-step fermentation from raw corn starch

“…Isoamylase (i.e., IA, EC 3.2.1.9, encoded by amyX gene) and pullulanase (i.e.,Pul,EC 3.2.1.41, encoded by pulA gene) specifically cleave the α-1,6-glycosidic linkages (Bi et al 2020;Ghosh et al 2020). Glucoamylase (i.e., GA, EC 3.2.1.3, encoded by glaA gene) not only cleaves the α-1,4-glycosidic linkages from non-reducing end but also slowly cuts the α-1,6-glycosidic linkages, thus producing glucose as the end product (Ghani et al 2013;Karim et al 2019;Lincoln et al 2019). It should be noted that the above mentioned amylolytic enzymes show the optimally active at weak acidity (i.e., pH 4.5 ~ 7.0) and high temperature (i.e., > 50 °C) (Guzmanmaldonado and Paredeslopez 1995).…”