Journal of South China University of Technology (Natural Science Edition) ›› 2022, Vol. 50 ›› Issue (1): 132-142.doi: 10.12141/j.issn.1000-565X.210145

Special Issue: 2022年生物工程

• Biological Engineering • Previous Articles     Next Articles

Heterologous Expression of Cutinase and Degradation of Polyethylene Terephthalate with Ultrasound

LOU Wenyong WU Youquan GUO Zewang ZONG Minhua   

  1. School of Food Science and Engineering,South China University of Technology,Guangzhou 510640,Guangdong,China
  • Received:2021-03-18 Revised:2021-06-10 Online:2022-01-25 Published:2022-01-03
  • Contact: 娄文勇(1975-),男,教授,博士生导师,主要从事应用生物催化研究。 E-mail:wylou@scut.edu.cn
  • About author:娄文勇(1975-),男,教授,博士生导师,主要从事应用生物催化研究。
  • Supported by:
    Supported by the National Natural Science Foundation of China(21676104,21878105),the National Key Research and Development Program of China(2018YFC1603400,2018YFC1602100) and the Key Research and Development Program of Guangdong Province(2019B020213001)

Abstract: Polyethylene terephthalate (PET) is a typical linear thermoplastic resin material,which is widely used in manufacture of fiber,film and beverage bottle.While PET brings many conveniences to human life,its property of difficult degradation has caused serious white pollution and threatened the global environment.In this research,the cutinase LCC was successfully expressed with Escherichia coli BL21 (DE3),and the optimized condition in E.coli BL21(DE3) expression is:inducer isopropyl-β-D-thiogalactopyranoside(IPTG)=0.2 mmol/L,induction temperature =16 ℃,and induction time =20 h.After protein purification process via Ni-affinity chromatography,the enzyme activity of LCC is 320 U/mg.Subsequently,the purified LCC was used to degrade PET with ultrasound,and their reaction conditions were optimized.Under the conditions of reaction temperature= 65℃,pH =8.0,ultrasonic power = 25 W,ultrasonic time=5 h,Km is the smallest and Vmax is the largest,the degradation efficiency of LCC with ultrasound is 1.21 times higher than that of untreated,and the product terephthalic acid concentration of (TPA) increases from 0.72 g/L to 0.87 g/L.According to the scanning electron microscopy (SEM),the degradation of PET catalyzed by LCC can be accelerated by ultrasound.The circular dichroism (CD) spectroscopy and fourier transform infrared (FT-IR) spectroscopy show that the α-helix and β-sheets of LCC secondary structure are possibly transformed into random coils with ultrasound.Thus,the degradation of PET catalyzed by LCC can be improved by ultrasonic process.This study provides some theoretical basis for the ultrasonic accelerated degradation reaction and opens up a new idea for the biodegradation of PET.

Key words: polyethylene terephthalate, ultrasound, biodegradation, cutinase, secondary structure

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