Efficient elimination and detection of phenolic compounds in juice using laccase mimicking nanozymes

Residual phenols in the juice can cause turbidity and affect its sensory quality.Laccase is used to remove phenolic compounds from fruit juice s.In order to overcome the shortcomings of natural laccase instability and high cost,in this work,we prepared a laccase mimic enzyme based on copper ion and adenosine monophosphate(AMP-Cu nanozymes).At the same mass concentration(1 mg·ml^(-1)), the catalytic activity of the nanozyme is about 15 times that of laccase.The AMP-Cu nanozymes had a higher V_(max) and a lower Km than laccase.The laccase mimic enzyme had a good stability under the condition of 30-90 ℃ and pH> 6.It also maintained high catalytic activity at high salt concentrations and 9 days storage time.Furthermore,the AMP-Cu nanozyme s maintained an initial catalytic activity of about 80% after six consecutive cycles of reaction.The linear range of detection of phenolic compounds by AMP-Cu nanozymes was 0.1-100 μmol·L^(-1) with a detection limit of 0.033 μmol·L^(-1).The phenol removal rate of AMP-Cu nanozymes was much higher than that of laccase under different reaction times.When the reaction was performed for 5 h,the phenol removal rate of the fruit juice by AMP-Cu nanozymes was about 65%.The efficient removal of phenolic compounds from different juices by AMP-Cu nanozymes indicate s that they have good application prospect in the food juice industry.

Application and optimization of solid-state fermentation process for enhancing polygalacturonase production by Penicillium expansum

Penicillium expansum 3.5425 was applied in solid-state fermentation(SSF)of agricultural wastes for polygalacturonase biosynthesis.Among various carbon additives,apple pomace was most suitable for the biosynthesis of polygalacturonase(1440.57 U/g).Optimization of medium parameters using rotational orthogonal design(ROD)experiment combined with optimal fermentation conditions resulted in a 2.72-fold increase in the polygalacturonase production.By using ammonium sulphate precipitation,ion-exchange and gel-permeation chromatography,the polygalacturonase produced by P.expansum 3.5425 was finally purified which had specific activity of 19269 U/mg and molecular weight of 30 kDa.The enzyme was remarkably active in the pH range of 3-5 and at 50℃,which makes it more acceptable in the industrial application.Besides,partially purified polygalacturonase(875.15 U/mL)was used for apple juice clarification and the clarity at 0.4 mL/kg was maximum,which reveals a great potential of polygalacturonase in food industry.