Abstract
Chitin is the second-most abundant bioresource and widely used in the food, agricultural, biomedicine, and other industries. However, under the mutual restriction of extraction cost and environmental protection, it is relatively difficult to prepare chitin from natural sources by pure separation. The aim of this study is to extract chitin from fresh crab shell waste by decalcification (DC) and deproteinization (DP) using glutamic acid and alkaline protease. The optimum technological conditions for DC and DP were as follows: (1) 5% (w/v) glutamic acid solution was used as decalcifying agent, the ratio of material to liquid was 1:10 (m/v), and the ash content in chitin was 0.83 ± 0.027% after decalcification at 75° C for 12 h. (2) Using alkaline protease as enzymatic hydrolyzer, 1500 U of alkaline protease was added per gram of crab shell. Under the conditions of material-liquid ratio of 1:10 (m/v) and pH value of hydrolysate of 9.0, N content in chitin was 6.63 ± 0.10% after 6 h of enzymatic hydrolysis at 55° C. And the extraction rate of chitin was 92.25 ± 0.51%. As a decalcifying agent, glutamic acid could be recycled with a recovery rate of 77.42 ± 2.16%. Calcium carbonate in crab shell was converted into calcium hydrogen phosphate by calcium glutamate, and protein into amino acids and polypeptides, which could be used as feed additives. The “glutamic acid-enzymolysis” for extracting chitin from crab shell is a relatively closed process, which has the advantages of mild reaction, greatly reducing the discharge of three wastes and high comprehensive utilization rate of raw materials.
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References
Liu, Y., Fang, G. H., Rong, S. H., Li, J. L., & Jiang, W. (2018). Advances in utilization of shrimp and crab shell. Food Safety and Quality Detection Technology, 9(3), 461–466.
Xu, Y., Mini, B., Reinhard, S., Stephan, L. G., Anne, S. U., Josef, W., & Claudia, G. (2013). Transformation of the matrix structure of shrimp shells during bacterial deproteination and demineralization. Microbial Cell Factories, 12(1), 90.
Iftekhar Shams, M., Nogi, M., Berglund, L. A., & Yano, H. (2012). The transparent crab: preparation and nanostructural implications for bioinspired optically transparent nanocomposites. Soft Matter, 8(5), 1369–1373.
Jia, R. C., & Nie, R. C. (2010). Extraction of chitin from lobster shell and preparation and properties of chitosan. Anhui Chemical Industry, 36(1), 41–43.
Vinayak, L. P., Karima, G., Tarek, R., & Saurabh, J. S. S. K. B. (2016). Novel biological and chemical methods of chitin extraction from crustacean waste using saline water. Journal of Chemical Technology&Biotechnology, 91(8), 2331–2339.
Choosit, H., Vitaliy, V. K., & Keshavan, N. (2016). Enzyme assisted extraction of chitin from shrimp shells (Litopenaeus vannamei). Journal of Chemical Technology&Biotechnology., 91(5), 1250–1256.
Hayes, M., Carney, B., Slater, J., & Brück, W. (2008). Mining marine shellfish wastes for bioactive molecules: chitin and chitosan-part a: extraction methods. Biotechnology Journal, 3(7), 871–877.
Liang, Y. J. (2013). Exyracting and applying chitin in antarctic krill shell. China: Dalian Polytechnic University.
Peng, Y. H., Wang, S., Li, C. P., Chen, B. T., & Huang, Y. L. (2017). Recycling of waste hydrochloric acid during chitin production process from Litopenaeus vannamei processing leftover. South China Fisheries Science, 13(1), 104–109.
Alabaraoye, E., Achilonu, M., & Hester, R. (2017). Biopolymer (chitin) from various marine seashell wastes: isolation and characterization. Journal of Polymers and the Environment, 26(6), 2207–2218.
Chou, Y. H., Doraiswamy, L. K., & Larson, M. A. (2001). Studies on the dissolution rate of sparingly soluble calcium citrate in water. Chemical Engineering Communications, 185(1), 223–236.
Rao, M. S., Nyein, K. A., Trung, T. S., & Stevens, W. F. (2007). Optimum parameters for production of chitin and chitosan from Squilla (S. empusa). Journal of Applied Polymer Science, 103(6), 3694–3700.
Kubantseva, N., & Hartel, R. W. (2002). Solubility of calcium lactate in aqueous solution. Food Reviews International, 18(2-3), 135–149.
Arbia, W., Adour, L., Amrane, A., & Lounici, H. (2013). Optimization of medium composition for enhanced chitin extraction from Parapenaeus longirostris by Lactobacillus helveticus using response surface methodology. Food Hydrocolloids, 31(2), 392–403.
Jung, W. J., Jo, G. H., Kuk, J. H., Kim, Y. J., Oh, K. T., & Park, R. D. (2007). Production of chitin from red crab shell waste by successive fermentation with Lactobacillus paracasei KCTC-3074 and Serratia marcescens FS-3. Carbohydrate Polymers, 68(4), 746–750.
Rai, A. K., Jini, R., Swapna, H. C., Sachindra, N. M., Bhaskar, N., & Baskaran, V. (2011). Application of native lactic acid bacteria (LAB) for fermentative recovery of lipids and proteins from fish processing wastes: bioactivities of fermentation products. Journal of Aquatic Food Product Technology, 20(1), 32–44.
Liu, L. P., Yin, Y. W., Sun, W. Q. & Ding, H. P. (2018). A method for extracting chitin from shrimp and crab shells by comprehensive treatment.CN 201810938635.4,2018-08-17, China.
Vijayalakshmi, P., & Sarvamangala, D. (2014). Production of L-glutamic acid by Arthrobacter globiformis MTCC 4299 using fruits of Mimusops elengi linn. International Journal of Applied Biology & Pharmaceutical Technology, 2(2), 167–173.
Zhu, Q. A. (2001). New technology for the one-step preparation of feed additive-calcium hydrophosphate. Yunnan Chemical Technology, 28(1), 16–18.
Synowiecki, J., & Al-Khateeb, N. A. (2003). Production, properties, and some new applications of chitin and its derivatives. Critical Reviews in Food Science and Nutrition, 43(2), 145–171.
Han, X. M., Wang, C. X., Yang, X., Wang, B., Sang, Y. X., & Sun, J. L. (2018). Study on preparation of calcium lactate and chitin from crab shell. Food Research and Development., 39(11), 65–70.
Aquatic industry standards of the People's Republic of China.SC/T3403-2018, China.
Determination of protein in foods. National standards of the People’s Republic of China.GB 5009.5-2016, China.
Aquatic industry standards of the People’s Republic of China.SC/T3403-2004, China.
Chen, Z. P., Wan, L. J., Wei, L. S., & Jiang, B. Q. (2014). Decalcification of bighead fish scales by citric acid leaching. Food & Machinery, 30(4), 151–154.
Tu, Z. C., Sha, X. M., Wang, H., Zhang, Q. T., Yao, S. H., Man, Z. Z., & Li, P. (2012). Studies on the decalcification of Aristichthys nobilis fish scale by acid. Food and Fermentation Industries, 38(2), 106–110.
Zhou, T., Duan, Z. Y., Zhang, H. J., Chen, J. X., & Mao, Z. G. (2004). Crystallization metastable zone properties of glutamic acid solution. Journal of Food Science and Biotechnology, 23(1), 62–65.
Hou, Y. K., Wang, S., Huang, K., Wang, W. J., Li, D., & Wang, J. Z. (2012). Optimization of walnut proteolysis process and analysis of antioxidant activity of enzymatic hydrolysate. Food and Fermentation Industries, 38(4), 99–103.
Liu, R. (2018). Comparison of Chitin Extraction from six common cultured insects. China: Hebei University.
Sun, B. B. (2016). Recombinant expression and application of enzymes in the resource utilization of shelis waste. China: Harbin Institute of Technology.
Liu, S., Sun, J., Yu, L., Zhang, C., Bi, J., Zhu, F., Qu, M. J., Jiang, C., & Yang, Q. L. (2012). Extraction and characterization of chitin from the beetle Holotrichia parallela Motschulsky. Molecules, 17(4), 4604–4611.
Funding
This work was supported by Zhejiang natural fund public welfare technology research program (LGN19C200009), Zhejiang province’s first-class discipline “bioengineering” open fund project (KF2019003), National college students innovation and entrepreneurship training program 2018 (Letter No.39[2018], 201810876005), Zhejiang province’s first-class discipline “bioengineering” student innovation program (CX2018024).
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Ding, H., Lv, L., Wang, Z. et al. Study on the “Glutamic Acid-Enzymolysis” Process for Extracting Chitin from Crab Shell Waste and its By-Product Recovery. Appl Biochem Biotechnol 190, 1074–1091 (2020). https://doi.org/10.1007/s12010-019-03139-2
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DOI: https://doi.org/10.1007/s12010-019-03139-2