COASTAL AQUACULTURE AUTHORITY Ministry of Agriculture, Government of India Farms permitted for culture SPF L. No Name of the Firm/Applicant & Address. Importancia y estudios de las comunidades microbianas en los recursos y productos pesqueros. Studies and importance of microbial communities. Conference Series LLC invites all the participants from all over the world to attend European Community Reference Laboratory for Crustacean Diseases leaflet 2008 Control and Prevention No effective vaccines for viruses within the Yellowhead complex. Production of High Viscosity Chitosan from Biologically Purified Chitin Isolated by Microbial Fermentation and Deproteinization. 2/13/2012 1 Aquaculture Global Outlook and Sustainability Travis Larkin Seafood Exchange Raleigh, NC. Coastal Aquaculture Authority. The Coastal Aquaculture Authority Act, 2005 (24 of 2005) enacted by the Parliment of India on 23 June 2005 provides for the. Centro de Investigaci. Correo Institucional; Red BIONNA; Licitaciones, Concursos y Subastas P Staphylococcus xylosus is a commensal of the skin of humans and animals and a ubiquitous bacterium naturally present in food. It is one of the major. Sharma Managing Director Mayank Aquaculture Private Limited. Surat (Gujarat State), India. E-mail: [email protected]. Production of High Viscosity Chitosan from Biologically Purified Chitin Isolated by Microbial Fermentation and Deproteinization. Department of Microbiology, Faculty of Science, King Mongkut’s University of Technology Thonburi, Pracha Uthit Road, Bangkok 1. Thailand. Received 1. September 2. 01. 3; Revised 1. December 2. 01. 3; Accepted 8 March 2. Published 2. 3 April 2. Copyright . This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The objective of this study was to produce high viscosity chitosan from shrimp chitin prepared by using a two- step biological treatment process: decalcification and deproteinization. Glucose was fermented with Lactobacillus pentosus L7 to lactic acid. At a p. H of , the calcium carbonate of the shells was solubilized in 4. The amounts of residual calcium in the form of ash () and crude protein () were further eliminated by the activity of proteolytic Bacillus thuringiensis SA. After decalcification and deproteinization of the shrimp shells, residual calcium and crude protein of shrimp chitin flakes were % and , respectively. Chitin was deacetylated with 5. Na. OH at 1. 21. After deacetylation, the chitosan had residual calcium, crude protein content, and degree of acetylation of , , and , respectively. The viscosity of chitosan prepared from chitin extracted by this two- step biological process was , whereas chitosan prepared from chemically processed chitin had a viscosity of , indicating that biologically purified chitin gave chitosan with a high quality. Introduction. The main unutilized biomass from the shrimp packaging and processing industries is heads and body carapaces, which constitute 4. About 2. 0–4. 0% of shrimp biowaste consists of chitin encrusted with calcium carbonate, protein, astaxanthin, and lipid residues . Shrimp biowaste is often treated in landfills or discarded in sea water, resulting in ecological problems in coastal areas, whereas a small part is used as a major component in chicken or fish feed, mixed with other agricultural raw materials . The exoskeletons of crustacean waste from the seafood industry are traditionally used to prepare commercial chitin and chitosan . Chitin can be converted to chitosan (. Commercial applications of chitosan are influenced by its viscosity in solution. The viscosity of chitosan depends strongly on the viscosity of the “preproduct” chitin . Chemical chitin extraction has a high efficiency for recovering purified chitin, but the process creates hazardous wastes which are harmful to human health and ecological systems . In addition, the chemical process has a negative effect on the intrinsic and physical properties of purified chitin, leading to a decrease in the viscosity of chitosan . Crude proteins and carotenoids in the extracting solution are useless after deproteinization and decalcification . Continued chitin production by chemical processes without development and utilization of novel technologies cannot solve the problem of achieving environmental sustainability . Currently, strictness in environmental protection has become a basic requirement for waste management in food and agricultural industries. To overcome the shortcomings of chemical chitin purification, several biotechnological techniques have been developed that are considered to be efficient alternative approaches for recovery of high quality chitin . The use of commercial crude enzymes to extract chitin influenced the cost production and also gave the low extraction efficacy . As a substitute for chemical and enzymatic processes, lactic acid fermentation combined with microbial deproteinization warrants further investigation because they are eco- friendly and positive procedures. The bacterial strain L. The objective of this work was to investigate a novel process for producing high viscosity chitosan from chitin isolated by two- step purification: the first step using the lactic acid bacterium L. Based on our knowledge, no prior study has reported the preparation of chitosan from biologically purified chitin prepared by using a combination of both strains. Materials and Methods 2. Raw Materials. Shells and heads of marine Pacific white shrimp (Litopenaeus vannamei) were obtained from a seafood wholesaler in Samut Sakhon province, Thailand. Both biowastes were packed into an ice box for transportation and stored frozen at . Before proximate analysis, both shrimp biowastes were washed thoroughly with tap water and dried in a hot- air oven at 1. For chitin and chitosan flake productions, shrimp shells were homogenized in a blender until small sized pieces (1. Microorganisms. The lactic acid bacterium L. Both strains were kept in a freezer at . Preparation of Shrimp Head Extract Solution (SHES)Shrimp head extract solution (SHES) was prepared by boiling shrimp heads in deionized water. Briefly, shrimp heads were mixed with deionized water at a ratio of 1 : 2 (5. SHES was obtained by filtering the solution through cotton to remove the sediment and then adding deionized water to obtain the initial liquid volume before heat extraction. Chemical Chitin Extraction. A mixture of 5. 00 g of wet shrimp shells and 2,2. L of 4% HCl was kept at room temperature for 4 hours to eliminate inorganic components in the shells. The decalcified shrimp shells were then separated and washed several times with tap water. Deproteinization was performed by adding the dried decalcified shrimp shells into 5% Na. OH at a ratio of 1 : 1. The mixture was shaken in a 9. Chitin flakes were obtained by washing the solid residue with tap water and drying overnight in an oven at 8. Biological Chitin Purification. A 2. 4- hour culture of L. Shrimp shell flakes (5. L of starter (1 : 1, w/v) followed by 1. L conical flask. Fermentation was performed at 3. The retentate was washed with tap water and dried in an oven overnight at 8. For deproteinization of the decalcified shrimp shells, 1. B. The mixture was incubated at 3. The chitin was washed and dried overnight in an oven at 8. Chitosan Production. Twenty m. L of 5. Deacetylation was performed at 1. Chitosan flakes were obtained by washing the solid residue with tap water until the p. H was neutral and then dried overnight in an oven at 8. Analytical Procedures. The p. H of the fermenting liquids was determined using a p. H meter (Metrohm, Riverview, FL, USA). Total titratable acid (TTA) was estimated by titration with 0. M Na. OH to neutralize all the total titratable protons. The endpoint was at p. H 8. 1. The calculation expresses the titratable acidity in terms of g/L of lactic acid. Reducing sugar was determined by the Somogyi- Nelson method . Moisture, protein, and ash contents were considered as the basis for determination of decalcification (DC) and deproteinization (DP) efficiencies. Moisture, fat, and ash contents of samples were determined by AOAC methods . Protein content was analyzed by a modified Biuret method, as described by Gornall et al. Briefly, 1. 00 mg dried materials were digested with 1. L 0. 5 M Na. OH for 4 hours at 4. DC and DP were calculated as percentages, according to Rao et al. The reaction was stopped by adding 6. The liquid was separated from the precipitate by centrifugation at 1. One unit of enzyme activity was defined as the amount which yielded an increase in of 0. The whiteness index (WI) was calculated based on the following . Chitin samples were dipped into 5. The chitin was then crushed into powder. N, N- dimethylacetamide (DMA) (9. Unilab; Ajax Finechem, NSW, Australia) containing 5% lithium chloride (Li. Cl) (9. 8. 0%, anhydrous; LOBA Chemie, Mumbai, India) was used to prepare 0. The mixture was stirred for 1. Chitosan solutions (1%, w/v) were obtained by dissolving chitosan powder in diluted acetic acid (1%, v/v). Each mixture was stirred for 2. A cone and plate geometry with a cone angle of 2. The shear rates ranged from 0. The experiments were conducted at 2. High, medium, and low viscosity chitosan represented viscosities of . Data Analysis. All experiments were run in triplicate. Experimental data were expressed as mean . Chemical Compositions of Raw Materials. Table 1 shows the composition of shrimp shells (abdominal part) and shrimp heads. The shrimp abdominal shells had less protein and lipid than shrimp heads; therefore, the abdominal part was the preferred raw material for chitin and chitosan preparations. The cost of the culture media for L. Shrimp heads are a readily available biowaste material, and SHES obtained by heat extraction process is cheap and easy to prepare. The SHES contained crude protein content of %, which could well support the growth of these bacteria. The residue after extraction still had some nutritional value (Table 2), which could be used for other applications such as animal feed production. Table 1: Chemical properties of shrimp shells and heads. Table 2: Chemical properties of shrimp head residue after heat extraction. Microorganisms Lactobacillus pentosus L7 was isolated from the Thai traditional fermented sausage, known as “Nham.” L. A protease- producing bacterium, B. Using the combination of both strains to decalcify and deproteinize shrimp shells, purified chitin with high quality could be obtained. The growth curves of L. The SHES could not support the growth of L. The modified MRS medium which used SHES to replace peptone, beef extract, and yeast extract could satisfactorily promote the growth of L. SHES plus 2% glucose could also promote the growth of L. SHES plus 2% glucose could save on the cost of inoculum medium; therefore, SHES plus 2% glucose medium was selected as the cultivation medium to prepare the inoculum for lactic acid fermentation of shrimp shells. When a 2. 4- hour culture of 0. B. The cell numbers of B. Two- Step Biological Chitin Purification of Shrimp Shells. Removal of protein and calcium from shrimp shells is critical step in chitin purification.
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