薏苡仁多菌发酵液的菌种优选及其抑制黑色素生成的作用研究

doi:10.19586/j.2095-2341.2022.0137

生物技术进展 ›› 2022, Vol. 12 ›› Issue (6): 929-936.DOI: 10.19586/j.2095-2341.2022.0137

薏苡仁多菌发酵液的菌种优选及其抑制黑色素生成的作用研究

毛新亮¹^,²(), 张卉³, 陈文杰¹, 陈艳武¹, 任红红², 余桂媛¹, 黄亚东³, 项琪³, 李晓敏¹^,²()

^1.广东完美生命健康科技研究院有限公司，广东中山 528451
^2.完美（广东）日用品有限公司，广东中山 528451
^3.广州暨南大学医药生物技术研究开发中心有限公司，广州 510000

收稿日期:2022-08-01 接受日期:2022-10-13 出版日期:2022-11-25 发布日期:2022-11-30
通讯作者: 李晓敏
作者简介:毛新亮 E-mail: 82998254@qq.com；
基金资助:
广州市科技计划项目(202103030003);校企合作产学研项目(E-SKY-YFZX-2020-012)

Study on Strain Optimization of Coix Seed Multi Lactobacillus Fermentation Broth and its Inhibition on Melanin Production

Xinliang MAO¹^,²(), Hui ZHANG³, Wenjie CHEN¹, Yanwu CHEN¹, Honghong REN², Guiyuan YU¹, Yadong HUANG³, Qi XIANG³, Xiaomin LI¹^,²()

^1.Perfect Life and Health Institute Co. ，Ltd. ，Guangdong Zhongshan 528451，China
^2.Perfect （Guangdong） Commodity Co. ，Ltd. ，Guangdong Zhongshan 528451，China
^3.Biopharmaceutical Research & Development Center，Jinan University，Guangzhou 510000，China

Received:2022-08-01 Accepted:2022-10-13 Online:2022-11-25 Published:2022-11-30
Contact: Xiaomin LI

摘要/Abstract

摘要：

以薏苡仁作为发酵基质，确定利于提高发酵液体外活性的较优乳酸菌种，并分析优势乳酸菌种薏苡仁发酵液对斑马鱼胚体黑色素生成的抑制作用。通过比较分析乳酸乳球菌（Lactococcus lactis）、嗜热链球菌（Streptococcus thermophilus）和保加利亚乳杆菌（Lactobacillus bulgaricus）3种单一乳酸菌和三者复合乳酸菌的薏苡仁发酵液的还原糖、总酚、游离氨基酸、蛋白、总酸和乳酸含量等理化指标及体外羟自由基清除能力和酪氨酸酶活抑制率确定较优发酵菌种，采用高通量测序测定发酵过程中微生物菌群结构；利用斑马鱼模型研究发酵液对黑色素生成的抑制作用。研究结果表明,采用乳酸乳球菌、嗜热链球菌和保加利亚乳杆菌3种乳酸菌复合发酵比单一乳酸菌发酵更具优势。使用以上菌种复合发酵薏苡仁过程中，乳酸乳球菌和嗜热链球菌为发酵前期优势菌群，发酵中后期则以保加利亚乳杆菌为优势菌群。经复合乳酸菌发酵后，薏苡仁发酵液的羟自由基清除率和酪氨酸酶活抑制率分别提高了20.82%和87.26%；斑马鱼模型实验结果表明，薏苡仁发酵液可以显著减少斑马鱼体表黑色素分布，当使用含量为2.0%时，对黑色素生成抑制率达到59.45%。研究结果为利用薏苡仁多菌发酵液开发为具美白肌肤性能的功能性新原料提供了科学数据支撑，并希望进一步推进薏苡产业的升级。

关键词: 薏苡仁, 乳酸菌, 发酵, 酪氨酸酶活, 黑色素

Abstract:

With coix seed as the fermentation substrate， the better lactic acid bacteria that can improve the activity in vitro of the fermentation liquid were screened， and the inhibitory effect of the dominant lactic acid bacteria coix seed fermentation broth on the melanin production of fish embryos was explored and analyzed. By comparing and analyzing the chemical indexes such as reducing sugar， total phenol， free amino acid， protein， total acid， lactic acid content， and hydroxyl radical scavenging ability and tyrosinase activity inhibition rate of coix seed fermentation broth of three kinds of single lactic acid bacteria Lactococcus lactis， Streptococcus thermophilus and Lactobacillus bulgaricus and three kinds of compound lactic acid bacteria of fermented coix seed with single or mixd Lactobacillus， the better fermentation strains were determined. High throughput sequencing was used to explore the microbial community structure and succession. The inhibitory effect of fermentation broth on melanin production was studied by zebrafish model. It was screened and determined that the compound fermentation of Lactococcus lactis， Streptococcus thermophilus and Lactobacillus bulgaricus had more advantages than that of single Lactobacillus. Lactococcus lactis and Streptococcusthermophilus were the dominant bacteria in the early stage of fermentation， and Lactobacillus bulgaricus was the dominant bacteria in the middle and late stage of fermentation. After fermentation with compound Lactobacillus， the hydroxyl radical scavenging rate and tyrosinase activity inhibition rate of fermented coix seed increased by 20.82% and 87.26%， respectively. The results of zebrafish model experiment showed that fermented coix seed could significantly reduce the distribution of melanin on zebrafish surface and when the content was 2%， the inhibition rate of melanin production could reach 59.45%. The research results can provide scientific data for the development of coix seed multi Lactobacillus fermentation broth as a new functional raw material with skin whitening properties， and promote the progress of coix seed industry.

Key words: Semen coicis, Lactobacillus, fermentation, tyrosinase activity, melanin

中图分类号:

TS275

毛新亮, 张卉, 陈文杰, 陈艳武, 任红红, 余桂媛, 黄亚东, 项琪, 李晓敏. 薏苡仁多菌发酵液的菌种优选及其抑制黑色素生成的作用研究[J]. 生物技术进展, 2022, 12(6): 929-936.

Xinliang MAO, Hui ZHANG, Wenjie CHEN, Yanwu CHEN, Honghong REN, Guiyuan YU, Yadong HUANG, Qi XIANG, Xiaomin LI. Study on Strain Optimization of Coix Seed Multi Lactobacillus Fermentation Broth and its Inhibition on Melanin Production[J]. Current Biotechnology, 2022, 12(6): 929-936.

图/表 4

表1 不同乳酸菌薏苡仁发酵液的主成分变化

Table 1 Changes of main substances in native and fermented coix seed fermented by different lactic acid bacteria

组别（n=3）	发酵前	发酵后
组别（n=3）	N	L.l	S.t	L.b	L.l+S.t+L.b
pH	5.82±0.02	3.90±0.01^*	3.48±0.00^*	3.25±0.03^*	3.21±0.01^*#
还原性糖/（mg·mL^-1）	62.89±3.79	58.94±1.88	59.65±2.40	65.76±6.23	61.91±1.72
游离氨基酸/（mg·mL^-1）	0.48±0.00	0.52±0.03	0.46±0.01	0.54±0.01	0.49±0.02
蛋白/（μg·mL^-1）	59.72±0.20	37.29±0.06^*	31.63±2.79^*	34.19±0.90^*	35.35±2.43^*
总酚/（μg·mL^-1）	57.58±1.24	62.40±0.38	55.58±1.36	60.34±0.72	61.34±2.47
总酸/（mg·mL^-1）	2.84±0.19	8.96±0.06^*	15.08±0.19^*	22.41±0.00^*	30.15±0.76^*##’&
乳酸/（mg·mL^-1）	ND	1.75±0.03^*	2.99±0.03^*	4.72±0.03^*	6.38±0.14^*##’&

图1 薏苡仁发酵前后对羟自由基清除率和酪氨酸酶活性抑制的影响注：*表示与阳性对照组相比，在P<0.05水平上具有统计学差异；#表示与发酵前（N）组相比，在P<0.05水平上具有统计学差异；#‘、&与#’分别表示L.l + S.t + L.b组与L.l组、S.t组与L.b组相比，在P<0.05水平上具有统计学意义。

Fig. 1 Inhibitory effects of native and fermented coix seed on hydroxyl radical scavenging and tyrosinase activity

图2 发酵过程总活菌数量和菌群相对丰度变化A：总活菌数量和发酸液pH变化；B：菌群相对丰度变化

Fig. 2 Changes in total number of viable bacteria and relative abundance of flora during fermentation

图3 薏苡仁发酵液抑制黑色素生成A：薏苡仁发酵液对斑马鱼胚胎形态学的影响；B：薏苡仁发酵液对鱼胚黑色素生成的抑制作用；C：发酵液对72 hpf时鱼胚酪氨酸酶活性的影响；*，**，***分别表示与对照组相比，差异在P<0.05、 P<0.01、 P<0.001水平上有统计学意义。

Fig. 3 Inhibition of melanin production by coix seed fermented coix seed

参考文献 32

1	LIU X, ZHANG X, RONG Y Z, et al.. Rapid determination of fat, protein and amino acid content in coix seed using near-infrared spectroscopy technique[J]. Food Anal. Methods, 2014, 8(2): 334-342.
2	JIE X X, GUO Z Y, PENG T Y, et al.. Assessment of the genetic diversity of different job's tears (coix lacryma-jobi l.) accessions and the active composition and anticancer effect of its seed oil[J/OL]. PLoS ONE, 2016, 11(4): e0153269 [2022-04-13]. .
3	ZHU F. Coix: chemical composition and health effects[J]. Trends Food Sci. Technol., 2017, 61: 160-175.
4	YIN H M, WANG S N, NIE S P, et al.. Coix polysaccharides: gut microbiota regulation and immunomodulatory[J]. Bioact. Carbohydr. Diet. Fibre, 2018, 16: 53-61.
5	杨志清,王硕,张世鲍,等.15种薏苡仁中多糖含量测定[J].亚太传统医药,2014,10(19):25-26.
6	ZHU R Y, XU X F, SHAN Q Y, et al.. Determination of differentiating markers in coicis semen from multi-sources based on structural similarity classification coupled with UPCC-Xevo G2-XS QTOF[J/OL]. Front. Pharmacol., 2020, 11: 549181[2022-04-13]. .
7	SOUSA C, HERRERA A R, BARRO F, et al.. The gluten-free diet: testing alternative cereals tolerated by celiac patients[J]. Nutrients, 2013, 5(10): 4250-4268.
8	张平,李晔,陈婵,等.灵芝菌株固体发酵对枇杷叶主要活性成分的影响[J].天然产物研究与开发,2016,28(2):242-246.
9	ZHANG Z H, FAN S T, HUANG D F, et al.. Effect of Lactobacillus plantarum NCU116 fermentation on asparagus officinalis polysaccharide: characterization, antioxidative, and immunoregulatory activities[J]. J. Agric. Food Chem., 2018, 66(41): 10703-10711.
10	ARORA S. Effect of germination and probiotic fermentation on nutrient profile of pearl millet based food blends[J]. British Food J., 2013, 113(4): 470-481.
11	RUSSO P, DE CHIARA M L V, CAPOZZI V, et al.. Lactobacillus plantarum strains for multifunctional oat-based foods[J]. LWT-Food Sci. Technol., 2015, 68: 288-294.
12	SONG A A, INLL, LIM S H, et al.. A review on Lactococcus lactis: from food to factory[J]. Microb. Cell Factories., 2017, 16(1): 55.
13	刘学云,于新,何嘉敏,等.九种益生菌之间的相互作用及协同共生机理[J].食品与发酵工业,2019,45(13):65-70.
14	LIU E, ZHENG H, SHI T, et al.. Relationship between Lactobacillus bulgaricus and Streptococcus thermophilus under whey conditions: focus on amino acid formation[J].Int. Dairy J., 2016, 56: 141-150.
15	李仲禧.苹果汁植物乳杆菌发酵对多酚化合物抗氧化活性的影响[D].上海:上海海洋大学,2019.
16	AVDEEF A, BERGER C M, BROWNELL C. pH-metric solubility. 2: correlation between the acid-base titration and the saturation shake-flask solubility-pH methods[J]. Pharm. Res., 2000, 17(1): 85-89.
17	郑梅,贾昌平.化妆品中10种α-羟基酸的高效液相色谱检测及质谱确证[J].中南药学,2022,20(4):919-923.
18	吴永祥,毕淑峰,姜薇,等.桑白皮多酚对B16细胞内黑色素生成的影响及其机制[J].中国药理学通报,2018,34(9): 1296-1301.
19	李朱昀,王未,李倩,等.百合珠芽与鳞茎营养成分及活性成分研究[J].生物技术进展,2016,6(5):336-340.
20	彭真汾,王威,谢倩,等.橄榄果实游离氨基酸测定方法的优化[J].食品工业科技,2017,38(22):263-267.
21	刘长姣,杨越越,王妮,等.茚三酮比色法测定黄秋葵氨基酸含量的不确定度评定[J].粮食与油脂, 2019,32(9):92-95.
22	AMANN R I, LUDWIG W, SCHLEIFER K H. Phylogenetic identification and in situ detection of individual microbial cells without cultivation[J]. Microbiol. Rev., 1995, 59(1): 143-169.
23	JIANG D, ZHENG M L, NIU D Z, et al.. Effects of steam explosion pretreatment and Lactobacillus buchneri inoculation on fungal community of unensiled and ensiled total mixed ration containing wheat straw during air exposure[J]. J. Appl. Microbiol., 2020, 128(3): 675-687.
24	樊英,于晓清,李乐,等.基于16S rRNA高通量测序分析大泷六线鱼表皮粘液及肠道内容物微生物多样性[J].生物技术进展,2021,11(1):79-90.
25	赵淑锐,杨源,郑美青,等.基于Fenton反应产生的羟自由基检测方法比较[J].实验技术与管理,2020,37(12):67-71.
26	李倩茹,夏珊,黄婷,等.建立酶标仪法检测沙棘的羟自由基清除活性[J].食品科技,2020,45(9):287-294.
27	李海馨,何婧芝,冯文亚,等.白芷美白活性部位初步研究[J].天然产物研究与开发,2018,30(3):425-428.
28	MAHDAVI A, MOHAMMADSADEGHI N, MOHAMMADI F, et al.. Evaluation of inhibitory effects of some novel phenolic derivatives on the mushroom tyrosinase activity: insights from spectroscopic analyses, molecular docking and in vitro assays[J/OL]. Food Chem., 2022, 387: 132938[2022-03-07]. .
29	DRIESSEN F M, KINGMA F, STADHOUDERS J. Evidence that Lactobacillus bulgaricus in yogurt is stimulated by carbon dioxide produced by Streptococcus thermophiles [J]. Netherland Milk Dairy J., 1982, 36: 135-144.
30	邓素芳,林忠宁,陆烝,等.薏苡产品开发与利用研究进展[J].粮食与饲料工业, 2016(6): 30-34.
31	王磊,阿地里江·阿不都拉,张富春,等.籽瓜瓤提取物对酪氨酸酶抑制作用及其成分分析[J].食品研究与开发, 2018, 39(19): 7-13.
32	CALAÇA G N, MACHADO S, WOHNRATH K, et al.. Simultaneous electroanalytical determination of depigmenting agents in skin-whitening products[J]. J. Electrochem. Soc., 2015, 162(12): 847-851.

[1]	李力群, 孙志康, 郝捷, 季嫱, 李选文, 吴晗, 吴娜, 郑超, 杨婧. 果胶酶生产及工业应用进展[J]. 生物技术进展, 2022, 12(4): 549-558.
[2]	张文静, 佟晔, 杨锡文, 曹彦金, 魏计东. 高产中性蛋白酶菌株的筛选、优化及中试放大[J]. 生物技术进展, 2022, 12(1): 112-119.
[3]	张春月, 金佳杨, 邱勇隽, 范立强, 赵黎明. 传统与未来的碰撞：食品发酵工程技术与应用进展[J]. 生物技术进展, 2021, 11(4): 418-429.
[4]	张兆昆,,周文学,李永丽,,胡建华,,刘占英,. 核黄素发酵菌种改造研究进展[J]. 生物技术进展, 2021, 11(1): 54-60.
[5]	王英燕,陈文娜,贾存江,房树标,秦斌,朱正乔. 发酵法生产制备番茄红素的工艺研究[J]. 生物技术进展, 2020, 10(3): 320-327.
[6]	王杰明,石雅丽,刘安礼,刘占英,兰辉,郝健霞,姚梽鹏,张清翠. 生物技术生产丁酸的研究进展[J]. 生物技术进展, 2020, 10(1): 53-59.
[7]	李宇邦,吴军林,李曼莎. 微生物发酵处理药食同源植物研究进展[J]. 生物技术进展, 2019, 9(5): 461-466.
[8]	费凡,冯伟伟,茆广华,张伟杰,丁阳阳,赵婷,吴向阳. 金针菇菌渣和醋渣固态发酵的工艺研究[J]. 生物技术进展, 2019, 9(4): 422-429.
[9]	邓展瑞,贠建民,郭娟,牛耀星,李彦虎. 陇西腊肉加工过程中优势乳酸菌的分离及其发酵性能研究[J]. 生物技术进展, 2019, 9(2): 200-209.
[10]	倪楠,辛志宏,张丽静,王凤忠,李淑英. 全豆乳液体发酵制备纳豆激酶工艺优化[J]. 生物技术进展, 2019, 9(2): 210-215.
[11]	赵彭年,杨德玉,王加友,丁一凡,安鹏,王远. 一株亚硝化细菌的分离鉴定及其发酵工艺优化[J]. 生物技术进展, 2019, 9(1): 69-77.
[12]	李彦虎,牛耀星,刘小霞,郭娟,邓展瑞,贠建民. 浆水直投式增香酵母冻干菌剂制备工艺的优化[J]. 生物技术进展, 2018, 8(5): 441-449.
[13]	郑威,周迎红,刘金龙4. 拟青霉A20140822发酵生产虫草素工艺的响应面优化研究[J]. 生物技术进展, 2017, 7(6): 625-631.
[14]	赵倩彦,姚允聪,闫静,姬谦龙. 果实发酵液作为肥料的功效研究进展[J]. 生物技术进展, 2017, 7(2): 135-138.
[15]	李小康,代京莎,李诚斌,陈茹,邱晓颖. 云芝菌株的选育、发酵和多糖提取研究进展[J]. 生物技术进展, 2017, 7(1): 20-24.

薏苡仁多菌发酵液的菌种优选及其抑制黑色素生成的作用研究

Study on Strain Optimization of Coix Seed Multi Lactobacillus Fermentation Broth and its Inhibition on Melanin Production

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 4

参考文献 32

相关文章 15

编辑推荐

Metrics

本文评价