Fish Gelatin Modification Using Prokaryotic Expression and Cell Surface Display Proline Hydroxylases

doi:10.13523/j.cb.2305027

China Biotechnology

2023, Vol. 43

Issue (11): 8-15 DOI: 10.13523/j.cb.2305027

Fish Gelatin Modification Using Prokaryotic Expression and Cell Surface Display Proline Hydroxylases

HUANG Ming-zhu^1,^2,³,SHEN Qi-chang^2,³,QIN Chun-yan^2,³,XU Yang³,WEI Yi-ran³,CHEN Xue-lan^1,^2,^3,^*(

)

1 National R&D Center of Freshwater Fish Processing, Jiangxi Normal University, Nanchang 330022, China
2 School of Life Science, Jiangxi Normal University, Nanchang 330022, China
3 School of Health, Jiangxi Normal University, Nanchang 330022, China

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Abstract

The poor gel properties of fish gelatin limit its commercial application. Enzyme-catalyzed modification of fish gelatin has great advantages such as environmental friendliness, safety and high efficiency. However, there are few gelatin-modified enzymes reported at present, and most of them are covalent cross-linking enzymes, which easily make gelatin form thermal irreversible gel. In this study, two proline hydroxylases that increase the non-covalent action of collagen were cloned and expressed in prokaryotic cells. After purification, the fish gelatin was catalyzed respectively using two proline hydroxylases. The results show that the two enzymes have the effect of improving the gel strength and texture characteristics of fish gelatin. In addition, the surface display technology of Corynebacterium crenatum was studied. The two enzymes were displayed on the surface of C. crenatum, and an immobilized enzyme system with C. crenatum as the carrier was prepared for studying the modification effect of two immobilized enzyme systems on fish gelatin. This study has enriched the catalytic enzyme system of fish gel and provided a new idea for the catalytic modification of fish gelatin.

Key words： Surface display technology Fish gelatin Gel strength Texture analysis

Received: 19 May 2023 Published: 01 December 2023

ZTFLH:

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Corresponding Authors: *Xue-lan CHEN E-mail: xuelanchen162@163.com

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	HUANG Ming-zhu
	SHEN Qi-chang
	QIN Chun-yan
	XU Yang
	WEI Yi-ran
	CHEN Xue-lan

Cite this article:

HUANG Ming-zhu, SHEN Qi-chang, QIN Chun-yan, XU Yang, WEI Yi-ran, CHEN Xue-lan. Fish Gelatin Modification Using Prokaryotic Expression and Cell Surface Display Proline Hydroxylases. China Biotechnology, 2023, 43(11): 8-15.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2305027 OR https://manu60.magtech.com.cn/biotech/Y2023/V43/I11/8

Table 1 Primers used in this study

Fig.1 SDS-PAGE of recombination protein M is the protein marker, the arrow indicated the purpose band, lanes 3 and 4 are samples induced without IPTG as control, and lanes 1,2,5 and 6 are Pbp4h fermentation samples, purified Pbp4h, Bap4h fermentation samples, and purified Bap4h, respectively

Table 2 Kinetic parameter of enzymes

Fig.2 Effect of enzyme catalysis on gel strength

Table 3 Effect of enzymatic catalysis on textural properties

Fig.3 Construction of Corynebacterium crenatum surface display system A. PCR fusion fragment; M is DNA marker, lanes 1-7 are gfp, ncgl1337-gfp, ncgl1337 transmembrane region sequence, pgsA-gfp, pgsA, proH-gfp, proH B. Fluorescence microscope observation C.Fluorescence intensity changes during liquid culture D. Each OD fluorescence intensity of each growth period

Fig.4 Effect of cells display the enzyme system catalysis on gel strength

Table 4 Effect of cells display the enzyme system catalysis on textural properties


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