产胞外多糖植物乳杆菌的筛选及粗多糖的活性研究

doi:10.7506/spkx1002-6630-201613022

摘要/Abstract

摘要：

通过观察乳酸菌菌落拉丝状况并测定其胞外多糖（exopolysaccharide，EPS）的产量，筛选出1 株所产EPS黏性好、产量高的乳酸菌AR307，经鉴定为植物乳杆菌。为得到更多的胞外多糖，对植物乳杆菌AR307的发酵条件进行优化，确定其在发酵温度32 ℃、发酵时间20 h条件下的产糖量为389 mg/L。在体外实验中，所得胞外多糖具有抑制HT-29肿瘤细胞活性、降低血糖水平的作用。

关键词: 植物乳杆菌, 发酵条件, 胞外多糖

Abstract:

The present study describes the screening of lactic acid bacterial (LAB) strains with high exopolysaccharide
(EPS)-producing ability by observing whether they could form sticky colonies. One strain named AR307 was finally
obtained and identified as Lactobacillus plantarum. In order to enhance EPS yield, the fermentation conditions for producing
EPS by Lactobacillus plantarum AR307 were optimized. The yield of EPS produced by AR307 reached up to 389 mg/L
after 20 h fermentation at 32 ℃. In vitro assay showed that the EPS had good inhibitory activity against colon cancer HT-29
cells and possessed an antidiabetic effect.

Key words: Lactobacillus plantarum, fermentation conditions, exopolysaccharide (EPS)

中图分类号:

TS201.3

冯小婉，夏永军，王光强，艾连中. 产胞外多糖植物乳杆菌的筛选及粗多糖的活性研究[J]. 食品科学, doi: 10.7506/spkx1002-6630-201613022.

FENG Xiaowan, XIA Yongjun, WANG Guangqiang, AI Lianzhong. Screening of Exopolysaccharide (EPS)-Producing Lactobacillus plantarum and Bioactivity of the EPS[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201613022.

参考文献

[1] BECK B R, KIM D, JEON J. The effects of combined dietary

probiotics Lactococcus lactis BFE920 and Lactobacillus plantarum

FGL0001 on innate immunity and disease resistance in olive flounder

(Paralichthys olivaceus)[J]. Fish Shellfish Immunology, 2015, 42(1):

177-183. DOI:10.1016/j.f si.2014.10.035.

[2] DUARY R K, BHAUSAHE M A, BATISH V K, et al. Antiinflammatory

and immunomodulatory efficacy of indigenous probiotic

Lactobacillus plantarum LP91 in colitis mouse model[J]. Molecular

Biology Reports, 2012, 39(4): 4765-75. DOI:10.1007/s11033-011-

1269-1.

[3] LADDA B, THEPAREE T, CHIMCHANG, et al. In vitro modulation

of tumor necrosis factor α production in THP-1 cells by lactic acid

bacteria isolated from healthy human infants[J]. Anaerobe, 2015, 33:

109-116. DOI:10.1016/j.Anaerobe.2015.03.002.

[4] WANG S C, CHANG C K, CHAN S C, et al. Effects of lactic acid

bacteria isolated from fermented mustard on lowering cholesterol[J].

Asian Pacific Journal of Tropical Biomedicine, 2014, 4(7): 523-528.

DOI:10.12980/ apjtb.4.201414b5.

[5] LIU J G, HOU C W, LEE S Y, et al. Antioxidant effects and UVB

protective activity of Spirulina (Arthrospira platensis) products

fermented with lactic acid bacteria[J]. Process Biochemistry, 2011,

46(7): 1405-1410. DOI:10.1016/j.Procbio.2011.03.010.

[6] KUDA T, KAWAHARA M, NEMOTO M, et al. In vitro antioxidant

and anti-inflammation properties of lactic acid bacteria isolated

from fish intestines and fermented fish from the Sanriku Satoumi

region in Japan[J]. Food Research International, 2014, 64: 248-255.

DOI:10.1016/j.Foodres.2014.06.028.

[7] MENDE S, PETER M, BARTELS K, et al. Addition of purified

exopolysaccharide isolates from S. thermophilus to milk and their

impact on the rheology of acid gels[J]. Food Hydroeolloids, 2013, 32:

178-185. DOI:10.1016/j.Foodhyd.2012.12.011.

[8] SURAYOT U, WANG J, SEESURIYACHAN P, et al.

Exopolysaccharides from lactic acid bacteria: structural analysis,

molecular weight effect on immunomodulation[J]. International

Journal of Biological Macromolecules, 2014, 68: 233-240.

DOI:10.1016/j.Ijbiomac.2014.05.005.

[9] VUYST L D, VIN F D, VANINGELGEM F, et al. Recent

developments in t h e biosynthesis and applications of

heteropolysaccharides from lactic acid bacteria[J]. International Dariy

Journal, 2001, 11(9): 687-707. DOI:10.1016/S0958-6946(01)00114-5.

[10] ARSKOLD E, SVENSSON M, GRAGE H, et al. Environmental

influences on exopolysaccharide formation in Lactobacillus reuteri

ATCC 55730[J]. International Journal of Food Microbiology, 2007,

116: 159-167. DOI:10.1016/j.ijfoodmicro.2006.12.010.

[11] HAJ-MUSTAFA M, ABDI R, SHEIKH-ZEINODDIN M.

Statistical study on fermentation conditions in the optimization

of exopolysaccharide production by Lactobacillus rhamnosus

519 in skimmed mi lk base media[ J ] . Biocatalys is and

Agricultural Biotechnology, 2015, 4(4): 521-527. DOI:10.1016

j.Bcab.2015.08.013.

[12] ALHUDHUD M, HUMPHREYS P, LAWS A. Development

of a growth medium suitable for exopolysaccharide production

and structural characterization by Bifidobacterium animalis ssp.

lactis AD011[J]. Microbiological Methods, 2014, 100(1): 93-98.

DOI:10.1016/j.Mimet.2014.02.021.

[13] de ALMEIDA JÚNIOR W L G, FERRARI Í D S, de SOUZA J V,

et al. Characterization and evaluation of lactic acid bacteria isolated

from goat milk[J]. Food Control, 2015, 53: 96-103. DOI:10.1016/

j.Foodcont.2015.01.013.

[14] MASUKE T, MINAMI A, IWASAKI N, et al. Carbohydrate analysis

by a phenol-sulfuric acid method in microplate format[J]. Analytical

Biochemistry, 2005, 339(1): 69-72. DOI:10.1016/j.Ab.2004.12.001.

[15] 张洁, 徐桂花, 尤丽琴. 16S rDNA序列分析法鉴定乳酸菌[J].

农产品加工( 创新版) , 2 0 0 9 ( 4 ) : 4 7 - 4 9 . D O I : 1 0 . 3 9 6 9 /

j.issn.1671-9646.2009.04.015.

[16] AI L Z, ZHANG H, GUO B H, et al. Preparation, partial

characterization and bioactivity of exopolysaccharides from

Lactobacillus casei LC2W[J]. Carbohydrate Polymers, 2008, 74(3):

353-357. DOI:10.1016/j.Carbpol.2008.03.004.

[17] 马杉姗, 王世平, 连正兴. 藏灵菇嗜热链球菌胞外多糖对人结肠癌

细胞的增殖抑制[J]. 食品科学, 2008, 29(04): 405-408.

[18] SIVASHANMUGAM A T, CHATTERJEE T K. In vitro and in vivo

antidiabetic activity of Polyalthia longifolia (Sonner.) Thw. leaves[J].

Oriental Pharmacy and Experimental Medicine, 2013, 13(4): 289-300.

DOI:10.1007/s13596-013-0118-2.

[19] RUAS-MADIEDO P, de los REYES-GAVILÁN C G. Invited

review: methods for the screening, isolation, and characterization

of exopolysaccharides produced by lactic acid bacteria[J]. Journal

of Dairy Science, 2005, 88(3): 843-856. DOI:10.3168/jds.S0022-

0302(05)72750-8.

[20] 凌代文, 冬秀珠. 乳酸菌分类鉴定及实验方法[M]. 北京: 中国轻工

业出版社, 1998: 117-129.

[21] 涂宏钢, 李存瑞, 巫庆华, 等. 利用16S rRNA全长序列鉴定植物

乳杆菌[J]. 乳业科学与技术, 2004, 27(2): 57-60. DOI:10.3969/

j.issn.1671-5187.2004.02.004.

[22] 熊素玉, 姚新奎, 谭小海. 不同温度及pH条件对乳酸菌生长影响的

研究[J]. 新疆农业科学, 2006, 43(6): 533-538.

[23] SUBUDHI S, BISHT V, BATTA N, et al. Purification and

characterization of exopolysaccharide bioflocculantproduced by

heavy metal resistant Achromobacter xylosoxidans[J]. Carbohydrate

Polymers, 2016, 137: 441-451. DOI:10.1016/j.Carbpol.2015.10.066.

[24] 冯美琴, 邢家溧, 张琦. 植物乳杆菌胞外多糖发酵条件的优化[J]. 食

品科学, 2011, 32(23): 215-219.

[25] GASSM M, SCHMIDTAND K A, FRANK J F. Exopolysaccharide

production from whey lactose by fermentation with Lactobacillus

celbrueckill ssp. bulgaricus[J]. Journal of Food Science, 1997, 62(1):

171-173; 207. DOI:10.1111/j.1365-2621.1997.tb04393.x.

[26] LI D, LI J, ZHAO F. The influence of fermentation condition on

production and molecular mass of EPS produced by Streptococcus

thermophilus 05-34 in milk-based medium[J]. Food Chemistry, 2016,

197: 367-372. DOI:10.1016/j.Foodchem.2015.10.129.

[27] 陈若雯, 李里, 段家彩. 乳酸乳杆菌胞外多糖的发酵条件优化[J]. 食

品与生物技术学报, 2010, 29(5): 770-776.

[28] MCKELLAR R C, GEEST J V, CUI W. Influence of culture and

environmental conditions on the composition of exopolysaccharide

produced by Agrobacterium radiobacter[J]. Food Hydrocolloids, 2003,

17: 429-437. DOI:10.1016/ S0268-005X(03)00030-4.

[29] WANG J, ZHAO X, TIAN Z, et al. Characterization of an

exopolysaccharide produced by Lactobacillus plantarum YW11

isolated from Tibet Kefir[J]. Carbohydrate Polymers, 2015, 125: 16-25.

DOI:10.1016/j.Carbpol.2015.03.003.

[30] WANG K, LI W, RUI X. Characterization of a novel exopolysaccharide

with antitumor activity from Lactobacillus plantarum 70810[J].

International Journal of Biological Macromolecules, 2014, 63: 133-139.

DOI:10.1016/j.Ijbiomac.2013.10.036.

[31] LIU L, JIA J, ZENG G. Studies on immunoregulatory and anti-tumor

activities of a polysaccharide from Salvia miltiorrhiza Bunge[J].

Carbohydrate Polymers, 2013, 92(1): 479-483. DOI:10.1016/

j.Carbpol.2012.09.061.

[32] DILNA S V, SURYA H, ASWATHY R G, et al. Characterization of

an exopolysaccharide with potential health-benefit properties from a

probiotic Lactobacillus plantarum RJF4[J]. LWT-Food Science and

Technology, 2015, 64(2): 1179-1186. DOI:10.1016/ j.lwt.2015.07.040.