Characteristics, Protein Engineering, Heterologous Production, and Industrial Applications of Microbial Isoamylases

Abstract

Isoamylase (EC 3.2.1.68), belonging to glycoside hydrolase subfamily GH13_11, catalyzes the hydrolysis of the 𝜶-1,6-D-glucosidic linkages in amylopectin, glycogen, and derived oligosaccharides. The high debranching efficiency and transglycosylation activity enable its use in food, feed, detergent, and other industries. However, the low catalytic efficiency and insufficient stability of currently known isoamylases prevent their widespread applications. To meet the requirements set by specific applications, therefore, there is a continuing demand to obtain isoamylases with good stabilities and high expression levels by protein engineering and development of novel fermentation strategies. Herein, the microbial sources, biochemical properties, and distinct sequence and structural features of microbial isoamylases are summarized. Current developments in the protein engineering, heterologous expression, and industrial applications of isoamylases are also discussed. Finally, the future research perspectives of microbial isoamylases are provided.

Protein engineering for natural product biosynthesis: expanding diversity for therapeutic applications

“…They applied engineering approaches that may be used to make drugs out of more potent but short-acting cytokines. [66] Applications with various industrially important enzymes Dong et al [67] conducted a literature study that included protein engineering applications, including various industrially relevant enzymes (such as nitrilases, aldolases, and xylosidases). Nitrilases are necessary biotransformation enzymes that hydrolyze nitriles into carboxylic acids and ammonia.…”

“…To address the demands of specific applications, protein engineering and the design of novel fermentation methods have been employed to manufacture isoamylases with better stabilities and high expression levels. [67] Other new applications New types of proteins have recently been generated using combinatorial protein engineering approaches. These binding proteins of non-Ig origin are known as "affibody binding ligands."…”

“…Dong et al [67] conducted a literature study that included protein engineering applications, including various industrially relevant enzymes (such as nitrilases, aldolases, and xylosidases). Nitrilases are necessary biotransformation enzymes that hydrolyze nitriles into carboxylic acids and ammonia.…”

“…To address the demands of specific applications, protein engineering and the design of novel fermentation methods have been employed to manufacture isoamylases with better stabilities and high expression levels. [67]…”

Purification, properties, and characterization of amylase: a review

“…Interestingly, amylase was the earliest biocatalytic macromolecule discovered by humans. In 1833, Payen and Persoz first obtained biocatalytic macromolecules with the function of decomposing starch [1] ; In 1924, Kuhn divided these biocatalytic macromolecules into two categories, namely, α-amylase and β-amylase, which are respectively it works by releasing α-and β-type maltose [2] ; In subsequent studies, scholars also discovered γ-amylase [3] and isoamylase [4] .…”