CHARACTERIZATION OF PARTIALLY PURIFIED TYROSINASE ISOLATED FROM BITTER KOLA (GARCINIA KOLA)

Objective: Tyrosinase is a glycosylated, copper-containing oxidase that catalyzes the first two steps of mammalian melanogenesis as well as enzymatic browning events in damaged fruits during post-harvest handling and processing. Human skin hyperpigmentation and enzymatic browning in fruits are both undesirable. In this study, the properties and inhibitory effect of some compounds on bitter kola tyrosinase were investigated. Methods: Bitter kola tyrosinase was isolated and characterized using standard protocols. Partial purification was carried out on Sephadex G-100 loaded column chromatography. Results: Bitter kola tyrosinase was purified with a specific activity of 3.5 U/mg protein, purification fold of 2.4 and a yield of 34%. The optimum pH value was found to be 6.0 while the optimum temperature value for maximum enzyme activity was observed at 60°C. The enzyme was stable at 40oC for 20 minutes. Metals such as NaCl, KCl, MgCl2 and CaCl2 had inhibitory effect on the activity; though MgCl2 and CaCl2 had minimal effect. Also, EDTA, β-marcaptoethanol and glutathione greatly inhibited the enzyme activity at all the tested concentration. Conclusion: The properties of bitter kola tyrosinase compare very well with the tyrosinase from other sources. Also, the study was able to establish the inhibitory effect of some compounds and this could be applied in food processing industries. Peer Review History: Received: 2 November 2021; Revised: 11 December; Accepted: 25 December, Available online: 15 January 2022 Academic Editor: Dr. A.A. Mgbahurike, University of Port Harcourt, Nigeria, [email protected] UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency. Received file: Reviewer's Comments: Average Peer review marks at initial stage: 6.0/10 Average Peer review marks at publication stage: 7.5/10 Reviewers: Dr. Nazim Hussain, North East Frontier Technical University, Arunachal pradesh, India, [email protected] Ahmad Najib, Universitas Muslim Indonesia, Makassar, Indonesia, [email protected] Prof. Dr. Ali Gamal Ahmed Al-kaf, Sana'a university, Yemen, [email protected] Similar Articles: PHYTOCHEMICAL PURIFICATION OF ACTIVE CONSTITUENTS ISOLATED FROM ROOT OF THE MEDICINAL HERB, CARALLUMA QUADRANGULA

Exogenous Salicylic and Ascorbic Acids Delay Peel Enzymatic Browning and Improve Quality of Dried Figs Under Low-Temperature Storage

Dried figs undergo undesirable changes during storage, particularly browning reactions. An approach to this issue is using, separately, antioxidants such as ascorbic acid and salicylic acid as anti-enzymatic browning agents. The aforementioned chemical agents were tested at the concentrations 0.1, 0.2, and 0.3% during 6 weeks-storage at 4 °C, for dried figs sealed within polyethylene terephthalate bags. For both agents, the results demonstrated that a concentration of 0.3% lowered the browning index compared to the control by 55 and 54%, respectively. Compared to other concentrations, the same concentration suppressed polyphenol oxidase to 75 and 80%, respectively. A significant impact (p < 0.05), for both treatments, particularly at 0.3%, was obtained on dried figs quality by lowering the total phenols loss and antioxidant capacity loss during the storage period. The results of vibrational spectroscopy were able to confirm the same pattern of the polyphenols compared to those examined by UV-Visible spectrophotometry, revealing thus a decreasing absorbance. A similar tendency was revealed using integrated intensity around the phenols vibration within the region of 1175–940 cm-1, as the concentrations of anti-browning agents increased. Thus, pre-storage ascorbic and salicylic acids treatments at 0.3% on dried figs could be appropriate to delay enzymatic browning and quality loss and, therefore to extend their shelf-life.

Antioxidant activity of thymol essential oil and inhibition of polyphenol oxidase enzyme: a case study on the enzymatic browning of harvested longan fruit

Background Enzymatic browning and microbial decay are the primary concerns that limit the postharvest life of longan fruit. These factors can be effectively prevented by sulfur dioxide (SO2) fumigation; however, due to the safety and regulatory issues of SO2, other alternatives must be tested. In this study, antioxidant and antimicrobial activities of thymol were determined against the pericarp browning and decay of longan fruit. A simple, cost-effective method was designed for its controlled release. Thymol vapors were obtained from the slurry prepared from 5 g of thymol in 5 mL of distilled water in a 180-mL glass jar, hermetically sealed and allowed for 24 h to accumulate the vapors in the headspace. Fruits were packed in polyethylene packages and fumigated with thymol through a septum. Non-fumigated fruits served as control and all the packages were stored at 25 ± 2 °C for 8 days. Results Thymol significantly (P ≤ 0.05) retarded pericarp browning (BI), delayed the decay incidence (DI) and maintained high color values of longan pericarp. Thymol also retained high total phenolic (TPC) and total flavonoid (TFC) contents, inhibited polyphenol oxidase (PPO) and peroxidase (POD) activities than those in control. A high coefficient of correlation of PPO with BI (r = 0.86), L* (r = − 0.94), weight loss (r = 0.93), TPC (r = − 0.77), TFC (r = − 0.80), DI (r = 0.92) and many other quality and color parameters indicated the antioxidant efficacy of thymol. Longer shelf life of 8 days with good quality attributes was obtained in thymol-treated fruits than 6 days in control. Conclusion Thymol could be effectively used as a natural antioxidant for a wide range of fruits. Graphic abstract

Comparative Transcriptomic Analyses Provide Insights into the Enzymatic Browning Mechanism of Fresh-Cut Sand Pear Fruit

Pear (Pyrus spp.) is one of the most commonly consumed temperate fruits, having considerable economic and health importance. Fresh-cut or processed pear fruits are prone to browning because of the abundant phenolic compounds; however, little is known about the molecular mechanisms underlying enzymatic browning of fresh-cut sand pear fruit. In this study, fruits of two sand pear genotypes (low browning cultivar ‘Eli No.2′ and high browning cultivar ‘Weiningdahuangli’) were used to analyze the molecular mechanism of enzymatic browning by SMRT-seq and RNA-seq. The results generated 69,122 consensus isoforms, 21,336 new transcripts, 7105 alternative splicing events, and 254 long non-coding RNAs (lncRNAs). Furthermore, five genes related to enzymatic browning were predicted to be targets of six lncRNAs, and 9930 differentially expressed genes (DEGs) were identified between two different flesh browning cultivars. Meanwhile, most DEGs (e.g., PAL, 4CL, CAD, CCR, CHS, and LAR) involved in the phenylpropanoid biosynthesis pathway were up-regulated, and the expression of PPO and POD were highly expressed in the high-browning cultivar. Interestingly, the transcript level of PbrPPO4 (Pbr000321.4) was significantly higher than other PPO and POD genes, and a high level of total polyphenol and PPO activity were observed in the high browning cultivar. We found that the expression of lncRNA PB.156.1 was significantly positively correlated with the target gene PbrPPO4 (Pbr000321.4). The results suggest that PbrPPO4 might act as a major contributor and a key enzyme encoding gene in regulating fresh-cut sand pear fruit enzymatic browning; the expression of PbrPPO4 was probably regulated by lncRNA PB.156.1. Altogether, the transcriptomic and physiological analyses expand the knowledge of sand pear flesh enzymatic browning at the molecular level and provide a foundation for germplasm resources for molecular breeding of high polyphenol and low browning cultivars in sand pears.

Changes in Browning Degree and Reducibility of Polyphenols during Autoxidation and Enzymatic Oxidation

In the present study, the browning degree and reducing power of browning products of catechin (CT), epicatechin (EC), caffeic acid (CA), and chlorogenic acid (CGA) in autoxidation and enzymatic oxidation were investigated. Influencing factors were considered, such as pH, substrate species and composition, and eugenol. Results show that polyphenols’ autoxidation was intensified in an alkaline environment, but the reducing power was not improved. Products of enzymatic oxidation at a neutral pH have higher reducing power than autoxidation. In enzymatic oxidation, the browning degree of mixed substrates was higher than that of a single polyphenol. The reducing power of flavonoid mixed solution (CT and EC) was higher than those of phenolic acids’ (CA and CGA) in autoxidation and enzymatic oxidation. Eugenol activity studies have shown that eugenol could increase autoxidation browning but inhibit enzymatic browning. Activity test and molecular docking results show that eugenol could inhibit tyrosinase.