Nutritional Enhancement of Polysaccharides from Photosynthetic Bacteria HP1 on Ophiocephalusargus Meat
-
摘要:
目的 筛选产多糖光合菌,为开发廉价易制备的益生菌多糖及应用提供参考。 方法 从虾塘中筛选一株产多糖的光合菌HP1,进行生理生化鉴定和16S rDNA序列比对。通过醇沉的方式从发酵液中制备光合菌HP1粗多糖,拌喂乌鳢30 d,以黄芪多糖作为参照,分别对比各个试验组对乌鳢肌肉营养成分的影响。 结果 HP1为粪红假单胞菌Rhodopseudomonasfaecalis。HP1胞外多糖组对乌鳢肌肉pH值没有显著影响(P>0.05),但能显著降低肌肉失水率并促使肌纤维直径变细(P < 0.05)。拌喂HP1胞外多糖对乌鳢粗蛋白含量无显著影响(P>0.05),但对乌鳢肌肉中的氨基酸组成有显著影响(P < 0.05),分别提高乌鳢肌肉中的总氨基酸、必需氨基酸、呈味氨基酸含量8.7%、11%、9%。另外,拌喂HP1胞外多糖也分别提高了乌鳢肌肉的粗脂肪含量、二十碳五烯酸(EPA)和二十二碳六烯酸(DHA)总含量10.7%和12.8%,显著提高肌肉中n-3与n-6系列脂肪酸比值。 结论 拌喂HP1胞外多糖不仅提高乌鳢肌肉的食用口感,也提升了乌鳢肌肉的营养价值,其效果与黄芪多糖拌喂的效果相似。 Abstract:Objective Enhancement on the development and application of exopolysaccharides produced economically from photosynthetic bacteria fermentation was investigated. Method A strain of polysaccharide-producing photosynthetic bacteria, HP1, was isolated from a shrimp aquaculture pond. Crude polysaccharides were prepared from fermentation broth and precipitated by ethanol. Using astragalus polysaccharides for comparison, effects of inclusion of the polysaccharides in feed on nutrients in the meat of Ophiocephalus argus fed for 30 d were examined. Result The physiological and biochemical characteristics as well as 16S rDNA sequencing of HP1 identified it to be Rhodopseudomonas faecalis. The polysaccharides produced from HP1 fermentation had no significant effect on muscle pH (P>0.05), but significantly decreased the drip loss and muscle fiber diameter of the fish after the feeding (P < 0.05). In the meat, the crude protein did not differ significantly by the diet change (P>0.05), but the amino acids did (P < 0.05). Compared with control without polysaccharide addition in the forage, the contents of total, essential, and delicious amino acids in the fish muscles increased by 8.7%, 11%, and 9%, respectively, crude fats by 10.7%, EPA+DHA by 12.8%, and the ratio of n-3/n-6. Conclusion It appeared that a diet included the exopolysaccharides not only improved the eating quality but also enhanced the nutritional value of O. argus with effects comparable to astragalus polysaccharides. -
表 1 乌鳢肌肉感官评价参考标准
Table 1. Reference standards for sensory evaluation on O. argus meat
项目
Items最好9~10分
Best (9-10 points)较好6~8分
Better (6-8 points)一般3~5分
Ordinary (3-5 points)最差0~2分
Worst (0-2 points)弹性Elasticity 肉质弹性好 肉质有弹性 肉质微有弹性 肉质杂乱无弹性 嫩度Tenderness 咀嚼时鲜嫩无残渣 咀嚼时鲜嫩,微有残渣 咀嚼时有粗糙感,有少量残渣 咀嚼时有粗糙感,有较多残渣 鲜味Fresh 鱼香味浓郁,鲜美 鱼香味较浓郁,较鲜 鱼香味较淡,微有鲜味 鱼香味极淡,无鲜味 甜味Sweet 甜味明显 微有甜味 甜味极淡 无甜味 表 2 菌株HP1生理生化特征
Table 2. Physiological and biochemical characteristics of HP1
项目
Items结果
Results最适温度
Optimum temperature34~37℃ 最适pH值
Optimum pH6.5~8.5 2%耐盐试验
2% Salt tolerance test+ 5%耐盐试验
5%Salt tolerance test+ 7%耐盐试验
7% Salt tolerance test― 明胶液化试验
Gelatin liquefaction test― 甲基红试验
Methyl red test― 过氧化氢酶试验
Catalase test+ 甘露醇
Mannitol+ 酪素
Casein― 淀粉
Starch― 柠檬酸盐
Citrate― 葡萄糖
Glucose+ 乙酸钠
Sodium acetate+ 丁酸钠
Sodium butyrate+ 丙酸盐
Propionate+ 表 3 菌株HP1的16S rDNA序列在GenBank的比对结果
Table 3. 16S rDNA sequence alignment of HP1 in comparison to GenBank database
编号
Serial numbers菌株
Strain序列覆盖率
Sequence coverage/%相似性
Similarity /%NR_024971.1 Rhodopseudomonas faecalis 100 99.44 HQ912770.1 Rhodopseudomonas sp. 100 99.09 HQ154127.1 Rhodopseudomonas faecalis 100 99.02 EU410078.1 Rhodopseudomonas faecalis 100 98.95 表 4 拌喂HP1胞外多糖后乌鳢肉质感官评价
Table 4. Sensory evaluation on meat of O. argus fed with HP1 exopolysaccharides
项目
ItemsC1 C2 C3 弹性Elasticity 5.0±0.2b 5.3±0.1b 6.2±0.3a 嫩度Tenderness 4.8±0.1b 6.3±0.5a 6.6±0.2a 鲜味Fresh 4.3±0.2b 5.7±0.1a 5.8±0.1a 甜味Sweet 4.2±0.3b 5.5±0.1a 5.9±0.3a 注:每组中同行平均值后不同字母表示差异显著(P<0.05)。表 5~8同。
Note:Data with different lowercase letters on the same line indicate significant difference at P<0.05. The same as Table 5-8.表 5 HP1胞外多糖对乌鳢肌肉理化性质影响
Table 5. Effect of HP1 exopolysaccharides on physicochemical properties of O. argus meat
项目
ItemsC1 C2 C3 肌纤维直径Diameter of muscle fiber/μm 75.3±2.1a 53.8±1.2b 56.7±2.8b 肌肉pH值pH of muscle 4.71±0.09 4.67±0.07 4.74±0.12 肌肉失水率Water loss rate of muscle /% 22.8±1.5a 18.4±1.1b 18.3±0.9b 表 6 HP1胞外多糖对乌鳢肌肉成分的影响
Table 6. Effect of HP1 exopolysaccharides on nutrient composition of O. argus muscles
项目
ItemsC1 C2 C3 肌肉粗脂肪Crude fat of muscle /% 7.52±0.35b 7.74±0.19b 8.48±0.53a 肌肉粗蛋白质Crude protein of muscle /% 88.69±6.83 88.20±5.27 87.12±5.15 单尾末重Weight of single one/g 647±23 651±18 655±19 表 7 HP1胞外多糖后对乌鳢肌肉氨基酸组成的影响
Table 7. Effect of HP1 exopolysaccharides on amino acids in O. argus muscles
(单位/%) 种类
Species氨基酸名称
Names of amino acidC1 C2 C3 甜味氨基酸Sweet amino acid 苏氨酸Thr 3.56±0.08b 3.92±0.03a 3.94±0.06a 脯氨酸Pro 3.49±0.04 3.51±0.11 3.60±0.18 赖氨酸Lys 7.19±0.07b 7.99±0.13a 7.93±0.15a 丝氨酸Ser 3.64±0.15 3.70±0.10 3.70±0.07 甘氨酸Gly 3.88±0.12b 4.30±0.09a 4.27±0.05a 丙氨酸Ala 4.38±0.05b 4.89±0.17a 4.87±0.12a 鲜味氨基酸Fresh amino acid 天冬氨酸Asp 8.13±0.12b 8.75±0.19a 8.88±0.24a 谷氨酸Glu 10.42±0.26b 11.69±0.16a 11.53±0.12a 含硫氨基酸Sulfur containing amino acid 胱氨酸Cys 0.94±0.15 1.03±0.18 1.08±0.10 蛋氨酸Met 2.07±0.18b 2.39±0.17a 2.38±0.07a 其他氨基酸Other amino acid 组氨酸His 2.25±0.04 2.28±0.12 2.38±0.08 缬氨酸Val 3.82±0.05b 4.17±0.09a 4.13±0.06a 异亮氨酸Iso 3.68±0.12b 4.03±0.05a 4.00±0.12a 亮氨酸Leu 6.13±0.17b 6.88±0.12a 6.85±0.14a 酪氨酸Tyr 2.45±0.05 2.52±0.09 2.48±0.06 苯丙氨酸Phe 3.45±0.13b 3.92±0.04a 3.93±0.08a 精氨酸Arg 4.78±0.05 4.83±0.21 4.84±0.16 氨基酸总和∑TAA 74.26±0.11b 80.8±0.12a 80.79±0.11a 人体必需氨基酸总和∑TEAAHB 29.9±0.10b 33.3±0.09a 33.16±0.08a 表 8 HP1胞外多糖后对乌鳢肌肉脂肪酸组成的影响
Table 8. Effect of HP1 exopolysaccharides on fatty acids in O. argus muscles
(单位/%) 脂肪酸
Fatty acidC1 C2 C3 十五烷酸C15:0 Pentadecylic acid C15:0 0.34±0.01 0.32±0.02 0.31±0.02 棕榈酸C16:0 Cetylic acid C16:0 23.23±0.27a 22.47±0.36b 22.09±0.48b 棕榈烯酸C16:1 Palmitoleic acid C16:1 3.70±0.05 3.55±0.18 3.56±0.13 十七烷酸C17:0 Daturic acid C17:0 0.34±0.02 0.32±0.02 0.31±0.03 硬脂酸C18:0 Octadecanoic acid C18:0 5.27±0.24 4.95±0.17 5.13±0.21 反式油酸C18:1T Trans oleic acid C18:1T 0.11±0.01 0.11±0.02 ― 顺式油酸C18:1 Cis oleic acid C18:1T 32.44±0.67a 31.61±0.85a 30.05±0.58b 顺式亚油酸C18:2 Cis linoleic acid C18:2 24.24±0.58 23.66±0.62 23.56±0.76 α亚麻酸C18:3N3 α-linolenic acid C18:3N3 0.22±0.02 0.32±0.01 0.31±0.01 γ亚麻酸C18:3n6 γ-linolenic acid C18:3n6 2.47±0.36 2.26±0.43 2.30±0.56 花生酸C20:0 Arachic acid C20:0 0.34±0.02 0.32±0.01 0.31±0.02 花生一烯酸C20:1 Cis-11-Eicosenoicacid C20:1 1.91±0.13a 1.94±0.09a 1.68±0.08b 二十一烷酸C21:0 n-heneicosanoic acid C21:0 0.56±0.01 0.65±0.05 0.52±0.02 γ-花生三烯酸C20:3N6 γ-Carbonium C20:3N6 0.11±0.01 0.11±0.01 ― 花生四烯酸C20:4N6 Arachidonic acid C20:4N6 0.56±0.04 0.43±0.08 0.52±0.01 γ-花生三烯酸C20:3N3 γ-Carbonium C20:3N3 0.22±0.01b 0.43±0.02a ― EPA C20:5 Eicosapentaenoicacid C20:5 1.01±0.25 0.75±0.12 1.05±0.23 DHA Docosahexaenoic acid C20:5 5.61±0.21b 4.95±0.28c 6.28±0.16a EPA+DHA 6.62±0.23b 5.70±0.19c 7.33±0.18a 饱和脂肪酸Saturated fatty acid 30.08±0.13a 29.03±0.16b 28.67±0.18c 单不饱和脂肪酸Monounsaturated fatty acid 38.16±0.21a 37.21±0.29b 35.29±0.26c 多不饱和脂肪酸Polyunsaturated fatty acid 26.93±0.32a 26.24±0.35b 26.17±0.23b 高不饱和脂肪酸Highly unsaturated fatty acid 7.51±0.1b 6.67±0.11c 7.85±0.13a n-3 7.07±0.12b 6.45±0.11c 7.64±0.13a n-6 3.14±0.14a 2.80±0.17b 2.82±0.29b n-3:n-6 2.25 2.31 2.71 -
[1] 李小勤, 胡斌, 冷向军, 等.VE对草鱼成鱼肌肉品质和抗氧化性能的影响[J].水生生物学报, 2009, 33(6):1132-1139. http://d.old.wanfangdata.com.cn/Periodical/ssswxb200906018LI X Q, HU B, LENG X J, et al. Effect of supplemental VE on meat quality and anti-oxidant capacity of adult grass carp[J]. ActaHydrobiologicaSinica, 2009, 33(6):1132-1139.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/ssswxb200906018 [2] 王清滨, 王秋举, 杨翼羽, 等.牛磺酸对投喂高脂饲料草鱼幼鱼生长、肌肉品质及抗氧化能力的影响[J].西北农林科技大学学报(自然科学版), 2015, 43(7):49-65. http://d.old.wanfangdata.com.cn/Periodical/xbnydxxb201507008WANG Q B, WANG Q J, YANG Y Y, et al. Effect of taurine on growth, muscle quality and antioxidant capacity of juvenile grass carp (Ctenopharyngodonidella) fed with high-fat diets[J]. Journal of Northwest A & F University (Natural Science Edition), 2015, 43(7):49-65.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/xbnydxxb201507008 [3] 王吉桥, 祁彩霞, 程爱香, 等.黄芪、熟地和山楂等中草药对黄颡鱼生长和消化的影响[J].水产学杂志2008, 21(1):34-41. http://d.old.wanfangdata.com.cn/Periodical/scxzz200801006WANG J Q, QI C X, CHENG A X, et al. Effects of astragalus, ripe and hawthorn on the growth and digestibility of juvenile yellow catfish[J]. Chinese Journal of Fisheries, 2008, 21(1):34-41.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/scxzz200801006 [4] 向枭, 陈建, 周兴华, 等.黄芪多糖对齐口裂腹鱼生长、体组成和免疫指标的影响[J].水生生物学报, 2011, 35(2):291-299. http://d.old.wanfangdata.com.cn/Periodical/ssswxb201102013XIANG X, CHEN J, ZHOU X H, et al. Effect of astragalus polysaccharides on growth, body composition and immune index in SchizothoraxPrenanti[J]. Acta Hydrobiologica Sinica, 2011, 35(2):291-299.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/ssswxb201102013 [5] 吴旋.四种中草药多糖对黄颡鱼生长、体成分及部分生理生化指标的影响[D].天津: 天津农学院, 2011. http://cdmd.cnki.com.cn/Article/CDMD-10061-1011191695.htmWU X. Effects of four kinds of Chinese herbal polysaccharides on the growth performance, body composition and part of physiological and biochemical indicator in yellow catfish (Pelteobagrus fulvidraco)[D]. Tianjin: Tianjin Agriculture University, 2011.(in Chinese) http://cdmd.cnki.com.cn/Article/CDMD-10061-1011191695.htm [6] 袁桂秀, 林晨, 江振友, 等.改良焦性没食子酸法培养厌氧菌[J].暨南大学学报, 1999, 20(4):24-25. http://www.cnki.com.cn/Article/CJFDTOTAL-JNDX199904006.htmYUAN G X, LIN C, JIANG Z Y, et al. Cultivating anaerobic bacteria by modifying pyrogallic acid method[J]. Journal of Jinan University (Medicine Edition), 1999, 20(4):24-25.(in Chinese) http://www.cnki.com.cn/Article/CJFDTOTAL-JNDX199904006.htm [7] 刘国红, 刘波, 陈峥, 等.一株产低温蛋白酶芽孢杆菌FJAT-24893的筛选与鉴定[J].福建农业学报, 2015, 30(10):954-957. http://www.fjnyxb.cn/CN/Y2015/V30/I10/954LIU G H, LIU B, CHEN Z, et al. Screening and phylogenetic analysis on a protease-producing Bacillus species[J]. Fujian Journal of Agricultural Sciences, 2015, 30(10):954-957.(in Chinese) http://www.fjnyxb.cn/CN/Y2015/V30/I10/954 [8] RAZA W, WU H S, SHEN Q.R. Use of response surface methodology to evaluate the effect of metal ions (Ca2+, Ni2+, Mn2+, Cu2+) on production of antifungal compounds by Paenibacilluspolymyxa[J]. Bioresource Technology, 2010, 101:1904-1912. doi: 10.1016/j.biortech.2009.10.029 [9] 郭振.改变水体盐度对吉富罗非鱼肉质的影响[D].天津: 天津农学院, 2014, 13-14. http://cdmd.cnki.com.cn/Article/CDMD-10061-1014225318.htmGUO Z. The effects of salinity changes on the muscle quality of Oreochromisniloticus[D]. Tianjin: Tianjin Agriculture University, 2014, 13-14.(in Chinese) http://cdmd.cnki.com.cn/Article/CDMD-10061-1014225318.htm [10] 郑永标, 江枝和, 李三暑, 等. 868发酵液提取物对甲鱼白细胞吞噬能力的影响[J].福建农业学报, 2001, 2(16):43-45. http://www.fjnyxb.cn/CN/Y2001/V/I2/43ZHENG Y B, JIANG Z H, LI S S, et al. Effect of 868 extraction on phagocytosis of leucocyte of Trionyxsinensis[J]. Fujian Journal of Agricultural Sciences, 2001, 2(16):43-45.(in Chinese) http://www.fjnyxb.cn/CN/Y2001/V/I2/43 [11] RAVI K P, PAPORI G, ABU T K, et al. Structural analysis and properties of dextran produced by Leuconostocmesenteroides NRRL B-640[J]. Carbohydrate Polymers, 2009, 76:30-35. doi: 10.1016/j.carbpol.2008.09.018 [12] LU Z Q, JIN M L, HUANG M, et al.Bioactivity of selenium-enriched exopolysaccharides produced by Enterobacter cloacaeZ0206 in broilers[J]. Carbohydrate Polymers, 2013, 96:131-136. doi: 10.1016/j.carbpol.2013.03.063 [13] REHFELDT C, FIEDLER I, DIETL G, et al. Myogenesis and postnatal skeletal cell growth as influenced by selection[J]. Livestock Production Science, 2000, 66(2):177-188. doi: 10.1016/S0301-6226(00)00225-6 [14] 石英, 冷向军, 李小勤.杜仲叶提取物对异育银鲫生长、血清非特异性免疫和肌肉品质的影响[J].浙江大学学报(农业与生命科学版), 2008, 34(2):200-206. http://d.old.wanfangdata.com.cn/Periodical/zjdxxb-nyysm200802013SHI Y, LENG X J, LI X Q, et al. Effect of eucommiaulmoidesoliver leaf extract on growth, serum non-specific immune response and meat quality of crucian carp[J]. Journal of Zhejiang University (Agric & Life Science Edition), 2008, 34(2):200-206.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/zjdxxb-nyysm200802013 [15] 舒琥, 周莹, 肖丹, 等.野生三斑石斑鱼肌肉营养成分与品质的评价[J].安徽农业科学, 2011, 39(21):12902-12905. http://d.old.wanfangdata.com.cn/Periodical/ahnykx201121096SHU H, ZHOU Y, XIAO D, et al. Evaluation of nutrient components and nutritive quality in muscle of wildEpinephelustrimaculatus[J]. Journal of Anhui Agricultural Science, 2011, 39(21):12902-12905.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/ahnykx201121096 [16] 邴旭文, 蔡宝玉, 王利平.中华倒刺鲃的肌肉营养成分与品质的评价[J].中国水产科学, 2005, 12(2):211-215. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=zgsckx200502018BING X W, CAI B Y, WANG L P. Evaluation of nutritive quality and nutritional components in Spinibarbussinensismuscle[J]. Journal of Fishery Sciences of China, 2005, 12(2):211-215.(in Chinese) http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=zgsckx200502018 [17] 胡兵, 刘军, 侯永清, 等.黄芪多糖对异育银鲫肌肉组成成分的影响[J].中国饲料, 2008(11):37-44. http://d.old.wanfangdata.com.cn/Periodical/zgsl200811017HU B, LIU J, HOU Y Q, et al. Effect of astragalus polysaccharides on nutritional ingredient in muscle of carassiusauratusgibelio[J]. China Feed, 2008(11):37-44.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/zgsl200811017 [18] 李卓佳, 林黑着, 郭志勋.中草药对斑节对虾生长、饲料利用和肌肉营养成分的影响[J].南方水产, 2007, 3(2):20-24. http://d.old.wanfangdata.com.cn/Periodical/nfsc200702004LI Z J, LIN H Z, GUO Z X. Effect of traditional Chinese medicines on growth, feed utilization and biochemical composition in muscle of juvenile shrimp Penaeusmonodon[J]. South China Fisheries Science, 2007, 3(2):20-24.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/nfsc200702004 [19] 王树海, 王文健, 汪雪峰, 等.黄芪多糖和小檗碱对3T3-L1脂肪细胞糖代谢及细胞分化的影响[J].中国中西医结合杂志, 2004, 24(10):926-928. http://d.old.wanfangdata.com.cn/Periodical/zxyjh200410017WANG S H, WANG W J, WANG X F, et al. Effect of astragalus polysaccharides and berberine on carbohydrate metabolism and cell differentiation in 3T3-L1 adipocytes[J]. Chinese Journal of Integrated Traditional and Western Medicine, 2004, 24(10):926-928.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/zxyjh200410017 [20] 王苗苗, 王海磊, 罗庆华.鳡鱼肌肉营养成分测定及评价[J].食品科学, 2014, 35(15):238-242. http://d.old.wanfangdata.com.cn/Periodical/spkx201415048WANG M M, WANG H L, LUO Q H, et al. Analysis and evaluation of nutrient components in muscle of yellow cheek carp[J]. Food Science, 2014, 35(15):238-242.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/spkx201415048