Structural and Functional Analysis of Pullulanase Type 1 (PulA) from Geobacillus thermopakistaniensis

Iqrar, Umber; Javaid, Hira; Ashraf, Naghmana; Ahmad, Aftab; Latief, Noreen; Shahid, Ahmad Ali; Ahmad, Waqar; Ijaz, Bushra

doi:10.1007/s12033-020-00255-x

Cited by 7 publications

(3 citation statements)

References 41 publications

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“…The signature motif of type I pullulanases, IYELHIRDEFS, being a part of (α/β) 8 TIM barrel was also conserved in CSR-VI of HL12Pul [9,22]. In addition, the consensus sequence YNWGYDP in CSR-VII, a highly conserved motif for all type I pullulanase enzymes, can be found in HL12Pul [9,12,23,24]. These therefore confirmed that HL12Pul was classified as type I pullulanase.…”

Section: Discussionsupporting

confidence: 65%

“…The biochemical characteristics of type I pullulanase from P. koreensis were described for the first time in this work. HL12Pul exhibited the highest identity to type I pullulanase from Priestia koreensis (99%) and was classified into subfamily 14 of glycoside hydrolase family 13 (GH13_14), which consists of type I pullulanases from a range of bacteria, such as Gram-positive bacteria Anaerobranca gottshalskii, Exiguobacterium acetylicum, Paenibacillus polymyxa, and Geobacillus thermopakistaniensis [9], while type I pullulanases belonging to GH13_12 and GH13_13 are generally identified from firmicutes, especially streptococci and plants, respectively [9,12]. The modular structure of HL12Pul consists of four domains, including (i) CBM68, (ii) CBM48, (iii) GH13 catalytic domain, and (iv) C-domain according to type I pullulanase from Anoxybacillus sp.…”

Section: Discussionmentioning

confidence: 99%

“…Among them, CBM20, CBM21, CBM41, CBM48, CBM53, and CBM68 have been identified in pullulanase type I. Moreover, undefined X domains are also located at N-terminal pullulanase, for instance, X25 and X45 domains [8,[12][13][14].…”

Section: Introductionmentioning

confidence: 99%

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In-Depth Characterization of Debranching Type I Pullulanase from Priestia koreensis HL12 as Potential Biocatalyst for Starch Saccharification and Modification

et al. 2022

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Pullulanase is an effective starch debranching enzyme widely used in starch saccharification and modification. In this work, the biochemical characteristics and potential application of a new type I pullulanase from Priestia koreensis HL12 (HL12Pul) were evaluated and reported for the first time. Through in-depth evolutionary analysis, HL12Pul was classified as type I pullulanase belonging to glycoside hydrolase family 13, subfamily 14 (GH13_14). HL12Pul comprises multi-domains architecture, including two carbohydrate-binding domains, CBM68 and CBM48, at the N-terminus, the TIM barrel structure of glycoside hydrolase family 13 (GH13) and C-domain. Based on sequence analysis and experimental cleavage profile, HL12Pul specifically hydrolyzes only α-1,6 glycosidic linkage-rich substrates. The enzyme optimally works at 40 °C, pH 6.0, with the maximum specific activity of 181.14 ± 3.55 U/mg protein and catalytic efficiency (kcat/Km) of 49.39 mL/mg·s toward pullulan. In addition, HL12Pul worked in synergy with raw starch-degrading α-amylase, promoting raw cassava starch hydrolysis and increasing the sugar yield by 2.9-fold in comparison to the α-amylase alone in a short reaction time. Furthermore, HL12Pul effectively produces type III-resistant starch (RSIII) from cassava starch with a production yield of 70%. These indicate that HL12Pul has the potential as a biocatalyst for starch saccharification and modification.

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