Ảnh hưởng của dịch chiết rong đỏ đến chất lượng tôm thẻ chân trắng (litopenaeus vannamei) bảo quản lạnh

Nội dung text: Ảnh hưởng của dịch chiết rong đỏ đến chất lượng tôm thẻ chân trắng (litopenaeus vannamei) bảo quản lạnh

1. KHOA H ỌC CƠNG NGHỆ P-ISSN 1859-3585 E-ISSN 2615-9619 THE EFFECT OF RED SEAWEED EXTRACT ON QUALITY OF WHITELEG SHRIMP (LITOPENAEUS VANNAMEI) DURING ICED STORAGE ẢNH HƯỞNG CỦA DỊCH CHIẾT RONG ĐỎ ĐẾN CHẤT LƯỢNG TƠM THẺ CHÂN TRẮNG (LITOPENAEUS VANNAMEI) BẢO QUẢN LẠNH Nguyen The Han1,*, Lam Thanh Ngoc1, Nguyen Thi Man1, Nguyen Thi Tuyet Nhu1, Dang Thi Thu Huong1, Khong Trung Thang2 1. INTRODUCTION ABSTRACT Melanin are pigments produced in a Tyrosinase plays an important role in the formation of melanin. Thus, the process of melanin well-known process called melanogenesis formation can be reduced through the inhibition of tyrosinase. The objective of this study is to (melanin biosynthesis), in the melanosomes investigate the tyrosinase inhibitory activity of different red seaweeds harvested in the coast of by melanocytes, which are distributed in the Khanh Hoa Province. All the tested extracts showed inhibitory effect on tyrosinase activity. The basal layer of the epidermis 1, 2. The color Gelidiella acerosa extract showed the most tyrosinase inhibitory activity with the IC50 value of expressed in skin, eye, and hair are the 3.04mg/mL. The effects of extraction conditions on tyrosinase inhibitory activity of G. acerosa were result of melanin, it serves an important investigated. Water was identified as the most effective solvent for the extraction. The suitable protective function against ultraviolet (UV) extraction conditions were found to be the solid to liquid ratio (g/mL) of 1/40, the extraction time of radiation 3, 4. More than 80% of the 60 min and the extraction temperature of 60C. The melanosis formation was significantly inhibited world’s population constitutes people with in whiteleg shrimp (Penaeus vannamei) treated with the G. Acerosa extract during storage in ice, white or yellow skin. Pheomelanin, which is compared with the control. Therefore, the red seaweed G. Acerosa could be used as a potential contained in these kinds of skins is much melanosis inhibitor in shrimp during refrigerated storage. less able to block UV energy and in fact may Keywords: Red seaweed, Gelidiella acerosa, tyrosinase inhibitor, melanosis, Litopenaeus vannamei synergize with UV photons to promote free radical formation, carcinogenesis in the skin TĨM TẮT 5, thereby resulting in aging. Tyrosinase đĩng vai trị quan trọng trong quá trình tổng hợp melanin. Do đĩ, quá trình tạo thành Tyrosinase, ubiquitously present in melanin cĩ thể được kiểm sốt thơng qua ức chế hoạt động của tyrosinase. Mục đích của nghiên cứu plants and animals, is the key enzyme in the này là đánh giá hoạt tính ức chế tyrosinase của dịch chiết một số lồi rong đỏ thu mẫu tại vùng biển synthesis of melanin that catalyzes Khánh Hịa. Kết quả nghiên cứu cho thấy, tất cả dịch chiết rong biển đều cĩ hoạt tính ức chế hydroxylation of L-tyrosine into L-Dopa tyrosinase. Dịch chiết rong Gelidiella acerosa cĩ hoạt tính ức chế cao nhất, với giá trị IC là 50 (monophenolase activity) and its oxidation 3,04mg/mL. Ảnh hưởng của điều kiện chiết đến hoạt tính ức chế tyrosinase của rong G. acerosa được to dopaquinone 6, 7. In addition to the nghiên cứu. Điều kiện chiết thích hợp được xác định như sau: Dung mơi chiết là nước, tỷ lệ nguyên formation of melanin pigment, tyrosinase liệu/dung mơi chiết (g/mL) là 1/40, thời gian chiết là 60 phút, nhiệt độ chiết là 60C. Hiện tượng tạo plays a key role in browning of fruits, thành điểm đốm đen (melanosis) của tơm thẻ chân trắng (Litopenaeus vannamei) bảo quản lạnh xử vegetables and sea foods 8. The lý bằng dịch chiết rong G. acerosa, được kiểm sốt một cách đáng kể so với mẫu đối chứng. Như vậy, rong đỏ G. acerosa là nguyên liệu tiềm năng sử dụng để hạn chế sự tạo thành điểm đốm đen trên tơm overexpression of tyrosinase may lead to trong quá trình bảo quản lạnh. dysregulation in melanogenesis which can trigger hyperpigmentation effects such as Từ khĩa: Rong đỏ, Gelidiella acerosa, chất ức chế tyrosinase, điểm đốm đen, Litopenaeus vannamei. melasma, freckles, age-related and 1Faculty of Food Technology, Nha Trang University chemical-spots etc. 9. In addition to this, 2Faculty of Mechanical Engineering, Nha Trang University tyrosinase has also been implicated in *Email: hannt@ntu.edu.vn playing a significant role in neurodegenerative disorders like Received: 26/5/2021 Parkinson’s disease 10, 11. In food, the Revised: 26/6/2021 quinones (dark, red, brown pigments) Accepted: 30/6/2021 formed by tyrosinase action cause 118 Tạp chí KHOA HỌC VÀ CƠNG NGHỆ ● Tập 57 - Số 4 (8/2021) Website:
2. P-ISSN 1859-3585 E-ISSN 2615-9619 SCIENCE - TECHNOLOGY enzymatic browning in plant-based foods and seafood, The samples were authenticated in place by a seaweed leading to deterioration during storage and commercial or expert (MSc. Do Anh Duy, Research Institute for Marine domestic processing 10, 12. Melanogenesis and Fisheries, Hai Phong city, Vietnam). Seaweed samples were enzymatic browning can be controlled in several ways, individually rinsed to remove impurities and air-dried in the including through the inhibition of tyrosinase gene shade. Dried seaweed samples were cut into small pieces, expression and inactivation of the related enzymes 13. vacuum packaged in PA bags and stored at 4°C until Many synthetic compounds have been demonstrated to analysis. whiteleg shrimps (Penaeus vannamei) with the size show inhibitory effect against tyrosinase enzyme and of 80 - 90 shrimp/kg were purchased from a local market in melanocyte on melanogenesis, such as kojic acid, mercury, Nha Trang city. These live shrimp were transported to the hydroquinone, and arbutin. However, they have been laboratory in plastic bags with aerated sea water within 30 associated with dangerous side effects in long-term use 7. min. l-3,4-Dihydroxyphenylalanine (l-DOPA) và tyrosinase Further research is therefore required to identify natural từ nấm (EC 1.14.18.1) were obtained from Sigma Aldrich (St. tyrosinase inhibitors that can be used as whitening agents Louis, MO, USA). All other reagents and solvents were of and antibrowning agents in the cosmetic and food analytical grade. industries respectively. 2.2. Seaweed extraction Marine seaweeds have been a great concern to To screen the tyrosinase inhibitory activitities, 10g of researchers; their multiple fascinating properties have each seaweed sample was extracted with 200mL 80% inspired a lot of studies on various species. Species from the aqueous methanol for 60 min at 60°C. The mixture was genus Laurencia (Rhodomelaceae, Ceramiales) belonging to filtered using Whatman No. 1 to obtain the crude extract. red algae, taxonomically classified as Rhodophyta, have been The solvent was removed using a rotary evaporator (IKA RV studied and established to possess important biological 10 control, Staufen, Germany) under vacuum pressure at activities such as antibacterial 14, antifungal 15 anti- 40°C. The obtained extracts were used to evaluate predatory 16, anticancer properties 17 and good source tyrosinase inhibitory activities. of natural antioxidant compounds, with defense systems The seaweed G. acerosa which showed potential potent enough to tolerate a wide range of stress-inducing tyrosinase inhibitory activity was selected to investigate the factors 18, 19. Red algae are primarily being considered in effect of solvent concentration on activities. Different this study because of their tyrosinase inhibitory properties solvent extractions (methanol, ethanol and water) were and their ability to retard oxidation that causes aging in used to extract under the same procedure as mentioned humans and enzymatic browning that causes discoloration above. The tyrosinase inhibitory activities were evaluated and aging in fruits and vegetables 20. to find out the suitable extraction solvent concentration. The tropical oceans of the world are enriched with the Then, seaweed was extracted using different conditions genus Laurencia, a significant producer of diterpenes, (temperature, time and seaweed/solvent ratio). halogenated sesquiterpenes, and acetogenins, making it the 2.3. Mushroom tyrosinase inhibitory assay world’s most chemically complex seaweed genus 21. Vietnam has a coastline of about 3260 km, with varying The inhibitory activity of mushroom tyrosinase was climatic conditions from the northern to southern parts of carried out using a spectrophotometric method with slight the country 22. These physical and climatological modifications of Chang et al. 24. In brief, 0.1mL of extract characteristics are suitable for the cultivation of seaweed. together with 2.2mL of phosphate buffer (pH 6.5), 0.1mL of The abundant algae floral estimated at nearly 1000 species substrate solution (2mM). The mixture was inculated at with about 827 species already identified 23, puts Vietnam 37C for 5 min. Then, 0.1mL of tyrosinase solution at an advantage. Seaweeds have long been cultivated in (100U/mL) was added and incubated at 37C for 30 min. Vietnam. Their biochemical properties have earned them a Absorbance was measured at 475nm after incubation. place in cuisines, traditional medicines, cosmetics, 2.4. Preparation of shrimp treated with seaweed extract pharmaceutical and more recently, ingredients for bio- The extract of G. acerosa with the highest tyrosinase industries. The objective of this study is to investigate the inhibitory activity was used for treatment of shrimp. Whole tyrosinase inhibitory activity of different red seaweeds shrimps were immersed in solution containing seaweed harvested in the coast of Khanh Hoa Province. The effect of extract at different concentrations (0; 0.78 and 1.59mg/mL, seaweed extract on the melanosis formation of whiteleg w/v) at 4°C for 30 min. Treated shrimps were drained on a shrimp during cold storage was also investigated. screen for 5 min at 4°C. Shrimps without seaweed extract 2. MATERIALS AND METHODS treatment were used as the control. The samples (25 shrimps) 2.1. Materials were placed on a polystyrene tray, covered with plastic wrap Five red seaweed species (Gelidiella acerosa, Gracilaria and stored in ice at 4°C. Samples were analyzed for salicornia, Acanthophora spicifera, Hypnea pannosa, melanosis and sensory every 3 days up to 12 days. Melanosis Kappaphycus alvarezii) were harvested during May to July, of shrimps was evaluated through visual inspection by ten 2018 at the coast of Nha Trang city, Khanh Hoa province. trained panelists using 10-point scoring test. Panelists were Website: Vol. 57 - No. 4 (Aug 2021) ● Journal of SCIENCE & TECHNOLOGY 119
3. KHOA H ỌC CƠNG NGHỆ P-ISSN 1859-3585 E-ISSN 2615-9619 asked to give the melanosis score (0 - 10), where 0 = absent; 3.2. Effect of extraction conditions tyrosinase inhibitory 2 = slight (up to 20% of shrimps’ surface affected); activity of G. acerosa 4 = moderate (20 - 40% of shrimps’ surface affected); The tyrosinase inhibitory activity of different solvent 6 = notable (40 - 60% of shrimps’ surface affected); 8 = severe (methanol, ethanol and water) extracts of G. acerosa was (60 - 80% of shrimps’ surface affected); 10 = extremely heavy investigated (Figure 1). All three solvents exhibited (80 -100% of shrimps’ surface affected) 25. different tyrosinase inhibitory activity. This indicates that 2.5. Statistical analyses the selection of an appropriate solvent for extraction of All analyses were performed in triplicate and all data are bioactive compounds from G. acerosa is critical to achieving expressed as the mean ± standard deviation (SD). The data optimal biological activity, particularly tyrosinase inhibitory were subjected to analysis of variance (ANOVA). Comparison activity. The aqueous extract, demonstrated the highest of means was carried out by the Duncan’s multiple range (55.49%) inhibitory activity while the ethanolic extract of test. Statistical analysis was performed using a SPSS package the species showed the least inhibitory activity (38.07%). (SPSS 16.0 for windows, SPSS Inc., Chicago, IL, USA). This shows that water is the preferred solvent for optimal G. 3. RESULT AND DISCUSSION acerosa tyrosinase inhibitory activity. Syad et al. 27 also recorded water as the best extraction solvent of G. acerosa. 3.1. Tyrosinase inhibitory activity of red seaweed extracts Water was therefore used as the solvent of extraction for The inhibitory effects of the five different species of red subsequent experiments. seaweed using mushroom tyrosinase were determined experimentally. Table 1 shows the tyrosinase inhibitory activity at different extract concentrations and the overall IC50 values which indicate 50% of the seaweeds’ potential in inhibiting tyrosinase activity. The IC50 values were different for the five species, varying from 3.04 to 6.53mg/mL. The tyrosinase inhibitory activity of the species decreased in the following order G. acerosa > H. pannosa > G. salicornia > K. alvarezii > A. spicifera. The differences in extract ability in inhibiting tyrosinase enzyme were majorly caused by differences in bioactive compounds contained in extracts originating from different species. The data demonstrate that G. acerosa had the highest tyrosinase inhibitory activity with IC50 value of 3.04mg/mL, it was therefore chosen for further investigation. Several studies investigated the Figure 1. Effect of different solvent extracts on tyrosinase inhibitory activity of phytoconstituents of G. acerosa, including flavonoids, G. acerosa. Different letters indicate statistically significant difference (p < 0.05) alkaloids, tannins, proteins, sulfated polysaccharides, sulfono glycolipid, sesquiterpenes, monoterpenes, phenols and various pharmacological activities 26. This is however, the first study investigating the tyrosinase inhibitory activity of the species. Table 1. Tyrosinase inhibitory activity of different red seaweeds Tyrosinase inhibitory activity (%) IC50 Seaweeds 2.0 2.5 3.0 3.5 4.0 value mg/mL mg/mL mg/mL mg/mL mg/mL (mg/mL) 33.01 44.66 50.74 63.94 60.03 3.04 G. acerosa ±1.22 ±2.11 ±1.54 ±2.09 ±0.44 ±0.01a 16.07 25.87 33.22 38.65 41.09 4.53 G. salicornia ±2.31 ±1.72 ±1.32 ±2.11 ±0.65 ±0.24b 25.58 31.50 34.08 35.83 36.98 6.53 A. spicifera ±3.19 ±1.99 ±1.51 ±4.91 ±1.03 ±0.47c 25.29 34.98 43.09 49.67 49.46 4.10 Figure 2. Effect of different extraction time on tyrosinase inhibitory activity of H. pannosa ±2.59 ±0.95 ±4.10 ±2.47 ±1.79 ±0.28b G. acerosa. Different letters indicate statistically significant difference (p < 0.05) 28.59 33.10 34.96 42.32 40.32 5.27 The effect of six different extraction time on tyrosinase K. alvarezii ±1.68 ±3.01 ±1.51 ±2.53 ±2.51 ±1.10bc inhibitory activity of aqueous extracts of G. acerosa was experimentally determined. Figure 2 shows that there were Different letters within the same column indicate significant difference significant differences among the percentage tyrosinase (p < 0.05) 120 Tạp chí KHOA HỌC VÀ CƠNG NGHỆ ● Tập 57 - Số 4 (8/2021) Website:
4. P-ISSN 1859-3585 E-ISSN 2615-9619 SCIENCE - TECHNOLOGY inhibitory activity of the extracts at the extraction times saturated with increasing solvent volume. Similar results on investigated (15, 30, 45, 60, 75, and 90 minutes). The the effect of the solid-liquid ratio on the extraction of tyrosinase inhibitory activity ranged from 16.29% to bioactive compounds from seaweeds were also reported 56.88%; extraction time of 75mins had the highest activity by Vázquez-Rodríguez et al. 30 and Topuz et al. 31. It at 56.88%. The aqueous extracts of G. acerosa exhibited a was concluded that the solid-liquid ratio of 1/40g/mL is time-dependent increase in tyrosinase inhibitory activity, suitable. however, after 75 minutes, increasing the extraction time did not improve the extract inhibitory activity of tyrosinase. Time is essential in optimizing energy requirements and cost during the extraction process. Since there was no significant difference between the inhibitory activity of the extract at 60 and 75 minutes, we can deduce that 60 minutes is the optimal extraction time for maximum tyrosinase inhibitory activity while minimizing energy costs. The use of different extraction temperature has significant effect (p < 0.05) on the tyrosinase inhibitory activity of aqueous extract of G. acerosa (Figure 3). The result showed that the inhibitory activity of the extract sharply increased when temperature increased from 30oC o to 60 C at 13.50% and 56.22%, respectively. However, o o increasing the temperature from 60 C to 90 C significantly Figure 4. Effect of different extraction ratio (w/v) on tyrosinase inhibitory reduced the inhibitory activity at 56.22% to 36.27%. This activity of G. acerosa. Different letters indicate statistically significant difference behavior can be attributed to the loss of biologically active (p < 0.05) compounds like polyphenols as a result of exposure to high 3.3. Effect of seaweed extract treatment on melanosis temperatures 28. It can be deduced from the result that of whiteleg shrimp during refrigerated storage 60oC is the most suitable extraction temperature for optimum inhibitory activity. The inhibition of melanosis in whiteleg shrimp using different concentrations of the G. acerosa extract was studied. Melanosis progression in the shrimp is shown in Figure 5. As expected, the melanosis score increased across all the groups with the control group having the highest mean scores (Figure 6). According to the panelists, none of the samples showed melanosis on day 0, melanin started to develop on the third day of storage which then began to increase significantly across the groups. Melanosis score of the shrimp was in descending order: control, 0.78mg/mL and 1.59mg/mL seaweed extract. Photographs of melanosis formation in whiteleg shrimp at different treatments for all the periods of refrigerated storage are shown in Figure 5. Samples in the photograph are representative of the same sample, which were evaluated Figure 3. Effect of different extraction temperature on tyrosinase inhibitory for melanosis score throughout the refrigerated storage. activity of G. acerosa. Different letters indicate statistically significant difference This study showed that immersing shrimp in seaweed (p < 0.05) extract inhibited post-harvest melanosis in shrimp; In the data presented, 1g of seaweed sample (G. additionally, the development of melanosis decreased as acerosa) was analyzed against varying volume (10 - 50mL) the concentration of the extract increased. Based on the of water to determine the solid/solvent ratio at which the finding, the seaweed extract concentration of 1.59mg/mL highest tyrosinase inhibitory activity could be obtained. was selected for preserving shrimp quality during The tyrosinase inhibitory activity increased with the solid- refrigerated storage. The inhibition of melanosis in shrimp liquid ratio increased until a maximum inhibitory activity immersed in plant extract has been reported in literature. reaches at 1/40g/mL (68.92%). The increase of the solid- Sharifian et al. 32 showed that 5% phlorotannins liquid ratio improves the rate of mass transfer, which results extracted from S. tenerimum inhibited melanosis in shrimp. in the increase in the mass transport driving force and the Llanto & Encarnacion 33 observed that the immersion of internal diffusion rate 29. However, the liquid-solid ratio shrimp in a 1.0% w/v solution of mushroom extract exhibited a slight decrease trend when the ratio was above significantly controlled melanosis in the treated shrimp 40mL/g, which was possible that the contact area was during ice storage and was comparable with the effects of Website: Vol. 57 - No. 4 (Aug 2021) ● Journal of SCIENCE & TECHNOLOGY 121
5. KHOA H ỌC CƠNG NGHỆ P-ISSN 1859-3585 E-ISSN 2615-9619 ascorbic acid and sodium sulfite treatments. Nirmal & determined as follows: the solid to liquid ratio (g/mL) of Benjakul 34 also reported a reduction in melanosis score 1/40, the extraction time of 60 min and the extraction in shrimp treated with tea extract. The retardation of temperature of 60C. The G. acerosa extract strongly melanosis in shrimp by the seaweed extract was most likely inhibited tyrosinase and melanoisis formation in whiteleg due to the high tyrosinase inhibitory activity of G. acerosa shrimp during 12 days of refrigerated storage. Thus, the G. (Table 1). acerosa extract could be used as a natural tyrosinase Control Seaweed extract Seaweed extract inhibitor for controlling melanoisis in shrimp during post (0.78mg/mL) (1.59mg/mL) storage. 0 day REFERENCE 1. D’mello S. A. N., Joseph W. R., Green T. N., Leung E. Y., During M. J., 3 Finlay G. J et al., 2016. Selected GRIN2A mutations in melanoma cause oncogenic day effects that can be modulated by extracellular glutamate. Cell Calcium, 60: 384- 395. 2. Zolghadri S., Bahrami A., Hassan Khan M. T., Munoz-Munoz J., Garcia- Molina F., Garcia-Canovas F., et al., 2019. A comprehensive review on tyrosinase 6 inhibitors. Journal of Enzyme Inhibition and Medicinal Chemistry, 34: 279-309. day 3. Slominski A., Tobin DJ., Shibahara S., Wortsman J., 2004. Melanin pigmentation in mammalian skin and its hormonal regulation. Physiological Reviews, 84: 1155-1228. 9 4. Wang G. H., Chen C. Y., Tsai T. H., Chen C. K., Cheng C. Y., Huang Y. H., et day al., 2017. Evaluation of tyrosinase inhibitory and antioxidant activities of Angelica dahurica root extracts for four different probiotic bacteria fermentations. Journal of Bioscience and Bioengineering, 123: 679-684. 5. Napolitano A., Panzella L., Monfrecola G., d’Ischia M., 2014. 12 Pheomelanin-induced oxidative stress: Bright and dark chemistry bridging red hair day phenotype and melanoma. Pigment Cell & Melanoma Research, 27: 721-733. 6. Cui H. X., Duan F. F., Jia S. S., Cheng F. R., Yuan K., 2018. Antioxidant and tyrosinase inhibitory activities of seed oils from torreya grandis Fort. ex Lindl. Figure 5. Melanois changes in whiteleg shrimps with and without treatment BioMed Research International, Article ID 5314320. of seaweed extract at different concentrations 7. Dolorosa M. T., Nurjanah P. S., Anwar E., Hidayat T., 2019. Tyrosinase inhibitory activity of Sargassum plagyophyllum and Eucheuma cottonii methanol extracts. IOP Conference Series: Earth and Environmental Science, 278. 8. Health A., Namjooyan F., Farasat M., Alishahi M., Jahangiri A., Mousavi H., 2019. The anti-melanogenesis activities of some selected brown macroalgae from northern coasts of the Persian Gulf. Brazilian Archives of Biology and Technology, 62. 9. Chang T. S., 2009. An updated review of tyrosinase inhibitors. International Journal of Molecular Sciences, 10: 2440-2475. 10. De Corradi I. F., De Souza E., Sande D., Takahashi J. A., 2018. Correlation between phenolic compounds contents, anti-tyrosinase and antioxidant activities of plant extracts. Chemical Engineering Transactions, 64: 109-114. Figure 6. The inhibition of melanosis in whiteleg shrimp using different 11. Hasegawa T., 2010. Tyrosinase-expressing neuronal cell line as in vitro concentrations of seaweed extract model of Parkinson’s disease. International Journal of Molecular Sciences, 11: 1082-1089. 4. CONCLUSIONS 12. Loizzo M. R., Tundis, R., Menichini, F., 2012. Natural and synthetic All red tested seaweed extracts showed inhibition tyrosinase inhibitors as antibrowning agents: An update. Comprehensive Reviews against mushroom tyrosinase inhibitory activity. Among in Food Science and Food Safety, 11:378-398. them, the G. acerosa extract showed the most tyrosinase inhibitory activity. The suitable extraction conditions for 13. Wang G. H., Chen C. Y., Tsai T. H., Chen C. K., Cheng C. Y., Huang Y. H., extraction of tyrosinase inhibitors from G. acerosa were et al., 2017. Evaluation of tyrosinase inhibitory and antioxidant activities of 122 Tạp chí KHOA HỌC VÀ CƠNG NGHỆ ● Tập 57 - Số 4 (8/2021) Website:
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