Recombinant Production of the Alpha-Amylase Enzyme from Tepidimonas fonticaldi in Lactococcus lactis

Project no. 31960 · Status: Kapanmış (Yayın bekleniyor) · 2023

Summary

<p>Microbial&nbsp;enzymes&nbsp;are&nbsp;widely&nbsp;used&nbsp;in&nbsp;industrial&nbsp;applications&nbsp;due&nbsp;to&nbsp;their&nbsp;high&nbsp;production&nbsp;yields&nbsp;and&nbsp;their&nbsp;ability&nbsp;to&nbsp;remain&nbsp;stable&nbsp;under&nbsp;a&nbsp;wide&nbsp;range&nbsp;of&nbsp;environmental&nbsp;conditions.&nbsp;Among&nbsp;these&nbsp;enzymes,&nbsp;α-amylase&nbsp;plays&nbsp;a&nbsp;significant&nbsp;role&nbsp;particularly&nbsp;in&nbsp;the&nbsp;food,&nbsp;detergent,&nbsp;and&nbsp;biofuel&nbsp;industries&nbsp;owing&nbsp;to&nbsp;its&nbsp;ability&nbsp;to&nbsp;hydrolyze&nbsp;starch.</p><p>In&nbsp;this&nbsp;study,&nbsp;the&nbsp;recombinant&nbsp;production&nbsp;of&nbsp;α-amylase&nbsp;derived&nbsp;from&nbsp;Tepidimonas&nbsp;fonticaldi&nbsp;was&nbsp;carried&nbsp;out&nbsp;in&nbsp;the&nbsp;Lactococcus&nbsp;lactis&nbsp;system,&nbsp;and&nbsp;the&nbsp;biochemical&nbsp;properties&nbsp;of&nbsp;the&nbsp;enzyme&nbsp;were&nbsp;investigated.&nbsp;The&nbsp;codon-optimized&nbsp;gene&nbsp;sequence&nbsp;was&nbsp;cloned&nbsp;into&nbsp;the&nbsp;pNZ8148&nbsp;vector,&nbsp;a&nbsp;recombinant&nbsp;system&nbsp;was&nbsp;established,&nbsp;and&nbsp;enzyme&nbsp;production&nbsp;was&nbsp;achieved.&nbsp;The&nbsp;activity&nbsp;of&nbsp;the&nbsp;extracellularly&nbsp;secreted&nbsp;enzyme&nbsp;was&nbsp;determined&nbsp;using&nbsp;the&nbsp;DNS&nbsp;method,&nbsp;and&nbsp;its&nbsp;characterization&nbsp;was&nbsp;performed&nbsp;in&nbsp;terms&nbsp;of&nbsp;pH,&nbsp;temperature,&nbsp;metal&nbsp;ions,&nbsp;detergents,&nbsp;and&nbsp;organic&nbsp;solvents.</p><p>The&nbsp;results&nbsp;demonstrated&nbsp;that&nbsp;the&nbsp;enzyme&nbsp;is&nbsp;active&nbsp;at&nbsp;high&nbsp;temperatures,&nbsp;stable&nbsp;over&nbsp;a&nbsp;wide&nbsp;pH&nbsp;range,&nbsp;and&nbsp;capable&nbsp;of&nbsp;retaining&nbsp;its&nbsp;activity&nbsp;in&nbsp;the&nbsp;presence&nbsp;of&nbsp;detergents.&nbsp;These&nbsp;findings&nbsp;indicate&nbsp;that&nbsp;the&nbsp;enzyme&nbsp;has&nbsp;potential&nbsp;for&nbsp;industrial&nbsp;applications.</p>

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