Effect of fermentation on hypoglycemic active ingredients in natural mulberry leaves

doi:10.16178/j.issn.0528-9017.20231233

Abstract

Abstract:

Fresh mulberry leaf sprouts were selected as the research object, and the traditional black tea fermentation process was used to improve the contents of hypoglycemic active ingredients in natural mulberry leaves through fermentation. The contents of different hypoglycemic active ingredients was used as a comprehensive evaluation index, and the different processing conditions affecting the fermentation effect were optimized through single factor test and response surface test, and the optimal fermentation process parameters were determined. The results showed that the contents of total alkaloids, total flavonoids and total polysaccharides in mulberry leaves increased by 27.92%, 28.71% and 20.10%, respectively, compared with natural mulberry leaves under the conditions of fermentation temperature of 40 ℃, fermentation time of 8.70 h and fermentation humidity of 85%. In conclusion, fermentation treatment can help to increase the content of effective hypoglycemic active ingredients in natural mulberry leaves, which can provide a reference for promoting the efficient utilization of natural mulberry leaves and increasing their added value.

Key words: natural mulberry leaves, fermentation conditions, response surface methodology, alkaloids, flavonoids, polysaccharides

CLC Number:

TS205.5

GUO Jieli, GAO Xin, YANG Limei, LIN Zhengshan, PAN Gebo. Effect of fermentation on hypoglycemic active ingredients in natural mulberry leaves[J]. Journal of Zhejiang Agricultural Sciences, 2025, 66(1): 189-196.

Figures/Tables 7

Table 1 Design and coding of experimental factor

水平	因素
水平	X₁/℃	X₂/h	X₃/%
-1	35	8	80
0	40	9	85
1	45	10	90

Fig.1 Influence of fermentation temperature on the contents of hypoglycemic ingredients in mulberry leaves

Fig.2 Influence of fermentation time on the contents of hypoglycemic ingredients in mulberry leaves

Fig.3 Influence of fermentation humidity on the contents of hypoglycemic ingredients in mulberry leaves

Table 2 Response surface parameter optimization results

序号	因素			响应值
序号	X₁/ ℃	X₂/ h	X₃/ %	Y₁/ (mg·g^-1)	Y₂/ (mg·g^-1)	Y₃/ (mg·g^-1)
1	0	-1	1	2.36±0.05	18.37±0.03	24.11±0.16
2	-1	0	-1	2.23±0.03	17.94±0.06	24.82±0.17
3	0	0	0	2.50±0.02	19.13±0.05	27.18±0.14
4	0	0	0	2.51±0.04	19.25±0.04	26.92±0.15
5	0	1	-1	2.34±0.06	18.32±0.07	22.13±0.12
6	-1	0	1	2.20±0.03	16.32±0.06	23.84±0.16
7	1	0	1	2.24±0.02	15.41±0.03	23.46±0.15
8	1	0	-1	2.28±0.01	16.11±0.05	23.74±0.14
9	0	1	1	2.26±0.04	17.19±0.06	23.37±0.18
10	0	-1	-1	2.39±0.05	18.68±0.04	25.76±0.15
11	1	1	0	2.21±0.03	15.07±0.07	23.13±0.13
12	0	0	0	2.51±0.06	19.23±0.08	27.00±0.18
13	-1	1	0	2.16±0.02	17.43±0.05	23.35±0.16
14	0	0	0	2.51±0.01	19.33±0.06	26.99±0.15
15	-1	-1	0	2.26±0.03	17.48±0.04	25.52±0.17
16	0	0	0	2.50±0.02	19.20±0.07	26.90±0.18
17	1	-1	0	2.31±0.05	16.85±0.08	24.37±0.15

Table 3 Analysis of variance in regression models

方差来源	Y₁/(mg·g^-1)			Y₂/(mg·g^-1)			Y₃/(mg·g^-1)
方差来源	F值	P值	显著性	F值	P值	显著性	F值	P值	显著性
模型	348.20	<0.000 1	^**	129.14	<0.000 1	^**	133.29	<0.000 1	^**
X₁	56.01	0.000 1	^**	153.89	<0.000 1	^**	27.12	0.001 2	^**
X₂	183.18	<0.000 1	^**	54.85	0.000 1	^**	206.48	<0.000 1	^**
X₃	47.97	0.000 2	^**	69.79	<0.000 1	^**	9.65	0.017 2	^*
X₁X₂	3.20×10^-3	0.956 7		28.67	0.001 1	^**	5.93	0.045 1	^*
X₁X₃	7.92×10^-2	0.786 5		7.06	0.032 6	^*	3.32	0.111 1
X₂X₃	10.30	0.014 9	^*	6.25	0.041 0	^*	57.25	0.000 1	^**
$X 1 2$	1 838.13	<0.000 1	^**	698.98	<0.000 1	^**	222.75	<0.000 1	^**
$X 2 2$	372.81	<0.000 1	^**	25.71	0.001 4	^**	264.08	<0.000 1	^**
$X 3 2$	381.79	<0.000 1	^**	68.30	<0.000 1	^**	309.86	<0.000 1	^**
失拟项	2.19	0.222 3	不显著	4.02	0.106 2	不显著	5.36	0.069 3	不显著
R²	0.997 8			0.994 0			0.994 2

Table 3 Analysis of variance in regression models

方差来源	Y₁/(mg·g^-1)			Y₂/(mg·g^-1)			Y₃/(mg·g^-1)
方差来源	F值	P值	显著性	F值	P值	显著性	F值	P值	显著性
模型	348.20	<0.000 1	^**	129.14	<0.000 1	^**	133.29	<0.000 1	^**
X₁	56.01	0.000 1	^**	153.89	<0.000 1	^**	27.12	0.001 2	^**
X₂	183.18	<0.000 1	^**	54.85	0.000 1	^**	206.48	<0.000 1	^**
X₃	47.97	0.000 2	^**	69.79	<0.000 1	^**	9.65	0.017 2	^*
X₁X₂	3.20×10^-3	0.956 7		28.67	0.001 1	^**	5.93	0.045 1	^*
X₁X₃	7.92×10^-2	0.786 5		7.06	0.032 6	^*	3.32	0.111 1
X₂X₃	10.30	0.014 9	^*	6.25	0.041 0	^*	57.25	0.000 1	^**
$X 1 2$	1 838.13	<0.000 1	^**	698.98	<0.000 1	^**	222.75	<0.000 1	^**
$X 2 2$	372.81	<0.000 1	^**	25.71	0.001 4	^**	264.08	<0.000 1	^**
$X 3 2$	381.79	<0.000 1	^**	68.30	<0.000 1	^**	309.86	<0.000 1	^**
失拟项	2.19	0.222 3	不显著	4.02	0.106 2	不显著	5.36	0.069 3	不显著
R²	0.997 8			0.994 0			0.994 2

Fig.4 Interaction effects among the various factors

References 27

[1]	XIA C L, TANG G H, GUO Y Q, et al. Mulberry Diels-Alder-type adducts from Morus alba as multi-targeted agents for Alzheimer’s disease[J]. Phytochemistry, 2019, 157: 82-91.
[2]	ZHANG R Y, ZHANG Q, ZHU S, et al. Mulberry leaf (Morus alba L.): a review of its potential influences in mechanisms of action on metabolic diseases[J]. Pharmacological Research, 2022, 175: 106029.
[3]	李来成, 张士凯, 许方舟, 等. 桑叶综合利用的研究进展[J]. 食品工业科技, 2022, 43(2): 397-404.
[4]	JAN B, PARVEEN R, ZAHIRUDDIN S, et al. Nutritional constituents of mulberry and their potential applications in food and pharmaceuticals: a review[J]. Saudi Journal of Biological Sciences, 2021, 28(7): 3909-3921.
[5]	何建新, 胡艳玲, 何汶珊, 等. 响应面-主成分分析法优化大曲发酵桑叶茶工艺[J]. 食品工业科技, 2021, 42(17): 113-119.
[6]	MA G Q, CHAI X Y, HOU G G, et al. Phytochemistry, bioactivities and future prospects of mulberry leaves: a review[J]. Food Chemistry, 2022, 372: 131335.
[7]	SUN C Z, WU W J, MA Y R, et al. Physicochemical, functional properties, and antioxidant activities of protein fractions obtained from mulberry (Morus atropurpurea Roxb.) leaf[J]. International Journal of Food Properties, 2017, 20(sup3): S3311-S3325.
[8]	ZHANG L W, SU S L, ZHU Y, et al. Mulberry leaf active components alleviate type 2 diabetes and its liver and kidney injury in db/db mice through insulin receptor and TGF-β/Smads signaling pathway[J]. Biomedicine & Pharmacotherapy, 2019, 112: 108675.
[9]	王婴. 桑叶活性部位对2型糖尿病microRNA调控效应研究[D]. 广州: 广州中医药大学, 2017.
[10]	杨清. 桑红茶工艺研究及其品质分析[D]. 重庆: 西南大学, 2010.
[11]	李莎, 王艳辉, 杨加虎, 等. 桑叶发酵综合利用的研究进展[J]. 北方蚕业, 2022, 43(2): 1-6, 11.
[12]	GUO N, ZHU Y W, JIANG Y W, et al. Improvement of flavonoid aglycone and biological activity of mulberry leaves by solid-state fermentation[J]. Industrial Crops and Products, 2020, 148: 112287.
[13]	SUN G X, LIANG Y, WANG J, et al. Fermentation process optimization of mulberry black tea by RSM[J]. Advanced Materials Research, 2013, 634/635/636/637/638: 1481-1485.
[14]	陈永丽, 高畅, 洛桑卓玛, 等. 米曲霉发酵桑叶茶的工艺优化[J]. 食品科技, 2020, 45(1): 114-121.
[15]	韩粮. 桑叶红茶加工工艺优化和开发[J]. 广东蚕业, 2018, 52(11): 7-9, 11.
[16]	孙国霞, 王俊, 龚敏, 等. 不同叶位桑叶及发酵温度对桑红茶中活性成分含量的影响[J]. 蚕业科学, 2011, 37(6): 1061-1066.
[17]	杨清, 徐立, 王俊, 等. 桑红茶发酵工艺条件优化及活性成分含量的动力学研究[J]. 蚕业科学, 2010, 36(2): 221-228.
[18]	郭娜. 桑叶和桑果活性成分提取与微生物转化研究[D]. 哈尔滨: 东北林业大学, 2021.
[19]	李凡, 裘雅渔, 钱文春, 等. 桑叶中总生物碱和1-脱氧野尻霉素的含量考察[J]. 中国药学杂志, 2008, 43(3): 176-179.
[20]	刘凡, 李平平, 廖森泰, 等. 98份不同桑树品种资源的桑叶总生物碱及1-脱氧野尻霉素含量测定[J]. 蚕业科学, 2012(2): 185-191.
[21]	高林森, 温吉更, 李亚丽, 等. 河北省不同地区、不同品种及生长时期桑叶中总黄酮含量分析[J]. 食品研究与开发, 2020, 41(2): 18-21.
[22]	WANG L H, SUN X Y, LI F, et al. Dynamic changes in phenolic compounds, colour and antioxidant activity of mulberry wine during alcoholic fermentation[J]. Journal of Functional Foods, 2015, 18: 254-265.
[23]	MA Q Q, SANTHANAM R K, XUE Z H, et al. Effect of different drying methods on the physicochemical properties and antioxidant activities of mulberry leaves polysaccharides[J]. International Journal of Biological Macromolecules, 2018, 119: 1137-1143.
[24]	杨斌旺, 刘平古, 刘龙飞, 等. 药桑叶多糖的脱蛋白工艺与抗氧化活性研究[J]. 食品研究与开发, 2020, 41(10): 156-160.
[25]	李旺, 韩蕴琦, 丁轲, 等. 发酵对蛋白桑1-脱氧野尻霉素提取量的影响[J]. 中国饲料, 2020(13): 45-49.
[26]	LIZARDO R C M, CHO H D, WON Y S, et al. Fermentation with mono- and mixed cultures of Lactobacillus plantarum and L. casei enhances the phytochemical content and biological activities of cherry silverberry (Elaeagnus multiflora Thunb.) fruit[J]. Journal of the Science of Food and Agriculture, 2020, 100(9): 3687-3696.
[27]	LIU G M, SUN J, HE X M, et al. Fermentation process optimization and chemical constituent analysis on Longan (Dimocarpus longan Lour.) wine[J]. Food Chemistry, 2018, 256: 268-279.