Aspergillus oryzae β-D-galactosidase immobilization on glutaraldehyde pre-activated amino-functionalized magnetic mesoporous silica: Performance, characteristics, and application in the preparation of sesaminol

Jinhong Gao; Lingli Zhang; Dongxin Zhao; Xin Lu; Qiang Sun; Heng Du; Hongyan Yang; Kui Lu

doi:10.1016/j.ijbiomac.2024.132101

Aspergillus oryzae β-D-galactosidase immobilization on glutaraldehyde pre-activated amino-functionalized magnetic mesoporous silica: Performance, characteristics, and application in the preparation of sesaminol

Int J Biol Macromol. 2024 May 10;270(Pt 1):132101. doi: 10.1016/j.ijbiomac.2024.132101. Online ahead of print.

Authors

Jinhong Gao¹, Lingli Zhang², Dongxin Zhao³, Xin Lu⁴, Qiang Sun⁴, Heng Du⁵, Hongyan Yang⁵, Kui Lu⁶

Affiliations

¹ College of Food Science and Engineering, Henan University of Technology, Zhengzhou, Henan 450044, China; Research Center for Agricultural and Sideline Products Processing, Henan Academy of Agricultural Sciences, Zhengzhou, Henan 450044, China.
² School of Chemical Engineering and Food Science, Zhengzhou University of Technology, Zhengzhou, Henan 450044, China.
³ School of Chemistry and Chemical Engineering, Henan University of Technology, Lianhua Road 100, Zhengzhou 450001, Henan Province, China.
⁴ Research Center for Agricultural and Sideline Products Processing, Henan Academy of Agricultural Sciences, Zhengzhou, Henan 450044, China.
⁵ College of Food Science and Engineering, Henan University of Technology, Zhengzhou, Henan 450044, China.
⁶ College of Food Science and Engineering, Henan University of Technology, Zhengzhou, Henan 450044, China; School of Chemical Engineering and Food Science, Zhengzhou University of Technology, Zhengzhou, Henan 450044, China. Electronic address: lukui126@126.com.

PMID: 38734354
DOI: 10.1016/j.ijbiomac.2024.132101

Abstract

Aspergillus oryzae β-D-galactosidase (β-Gal) efficiently hydrolyzes sesaminol triglucoside into sesaminol, which has higher biological activity. However, β-Gal is difficult to be separate from the reaction mixture and limited by stability. To resolve these problems, β-Gal was immobilized on amino-functionalized magnetic nanoparticles mesoporous silica pre-activated with glutaraldehyde (Fe₃O₄@mSiO₂-β-Gal), which was used for the first time to prepare sesaminol. Under the optimal conditions, the immobilization yield and recovered activity of β-Gal were 57.9 ± 0.3 % and 46.5 ± 0.9 %, and the enzymatic loading was 843 ± 21 U_enzyme/g_support. The construction of Fe₃O₄@mSiO₂-β-Gal was confirmed by various characterization methods, and the results indicated it was suitable for heterogeneous enzyme-catalyzed reactions. Fe₃O₄@mSiO₂-β-Gal was readily separable under magnetic action and displayed improved activity in extreme pH and temperature conditions. After 45 days of storage at 4 °C, the activity of Fe₃O₄@mSiO₂-β-Gal remained at 92.3 ± 2.8 %, which was 1.29 times than that of free enzyme, and its activity remained above 85 % after 10 cycles. Fe₃O₄@mSiO₂-β-Gal displayed higher affinity and catalytic efficiency. The half-life was 1.41 longer than free enzymes at 55.0 °C. Fe₃O₄@mSiO₂-β-Gal was employed as a catalyst to prepare sesaminol, achieving a 96.7 % conversion yield of sesaminol. The excellent stability and catalytic efficiency provide broad benefits and potential for biocatalytic industry applications.

Keywords: Aspergillus oryzae; Enzyme immobilization; Glutaraldehyde; Magnetic mesoporous silica; Sesaminol; β-D-galactosidase.