一株新型烟叶淀粉降解菌的筛选鉴定及其发酵条件优化

doi:10.3969/j.issn.1004-1524.20250222

浙江农业学报 ›› 2026, Vol. 38 ›› Issue (2): 206-216.DOI: 10.3969/j.issn.1004-1524.20250222

一株新型烟叶淀粉降解菌的筛选鉴定及其发酵条件优化

周航¹(), 孙梦瑶¹, 杨艾勇², 余洋洋¹, 裘垚¹, 杨永锋³, 陈红丽¹^,^*()

1.河南农业大学烟草学院, 河南郑州 450002
2.大理州烟草公司弥渡县分公司, 云南弥渡 675600
3.河南中烟工业有限责任公司技术中心, 河南郑州 450016

收稿日期:2025-03-19 出版日期:2026-02-25 发布日期:2026-03-24
作者简介:周航,主要从事烟草质量评价研究。E-mail:zhou813010@163.com
通讯作者: ^*陈红丽,E-mail: chenhonglili06@163.com
基金资助:
中国烟草总公司科技项目(110202202029)

Screening, identification and fermentation condition optimization of a novel starch-degrading bacterium isolated from tobacco leaves

ZHOU Hang¹(), SUN Mengyao¹, YANG Aiyong², YU Yangyang¹, QIU Yao¹, YANG Yongfeng³, CHEN Hongli¹^,^*()

1. College of Tobacco, Henan Agricultural University, Zhengzhou 450002, China
2. Midu County Branch of Dali Tobacco Company, Midu 675600, Yunnan, China
3. Technology Center of Henan China Tobacco Industry Co., Ltd., Zhengzhou 450016, China

Received:2025-03-19 Online:2026-02-25 Published:2026-03-24

摘要/Abstract

摘要：

为有效降低烟叶淀粉含量以提升品质,从烟叶表面筛选出可高效降解烟叶淀粉的菌株K1,经分子生物学鉴定,确定其为鲁戈斯芽孢杆菌(Bacillus rugosus)。基于单因素试验和响应面分析,探究最利于该菌株降解烟叶淀粉的发酵条件。结果表明:其最适碳源、氮源和无机盐分别为麦麸、豆粕和KH₂PO₄。经优化,其最适发酵条件为:麦麸13.6 g·L^-1,豆粕11.7 g·L^-1,KH₂PO₄ 12.5g·L^-1,pH值8.0,转速200 r·min^-1,发酵温度32 ℃。在此条件下,其淀粉酶活性达145.08 U·mL^-1,发酵后烟叶的淀粉含量下降49.43%,总糖和还原糖含量分别增加9.11%和11.05%。研究结果为降解烟叶淀粉、提高烟叶质量提供了优良的菌种资源,为烟叶淀粉降解菌的进一步开发利用奠定了基础。

关键词: 烟草, 淀粉降解菌, 菌株筛选, 菌株分离, 菌株鉴定, 培养基优化, 发酵条件

Abstract:

To effectively reduce the starch content in tobacco leaves and improve the quality, a strain K1 with high-efficiency starch-degrading capability was isolated from the surface of tobacco leaves. Molecular biological identification confirmed that the strain was Bacillus rugosus. Based on single-factor experiments and response surface methodology, the optimal fermentation conditions for the strain to degrade tobacco leaf starch were investigated. The results showed that the optimal carbon source, nitrogen source and inorganic salt were wheat bran, soybean meal and potassium dihydrogen phosphate (KH₂PO₄), respectively. After optimization, the optimal fermentation conditions were determined as follows: wheat bran at 13.6 g·L^-1, soybean meal at 11.7 g·L^-1, KH₂PO₄ at 12.5 g·L^-1, initial pH value of 8.0, rotation speed of 200 r·min^-1 and fermentation temperature of 32 ℃. Under these conditions, the amylase activity of the strain reached 145.08 U·mL^-1. After fermentation, the starch content in tobacco leaves decreased by 49.43%, while the contents of total sugar and reducing sugar increased by 9.11% and 11.05%, respectively. This study provides an excellent strain resource for degrading tobacco leaf starch and improving tobacco leaf quality, and lays a foundation for the further development and utilization of tobacco leaf starch-degrading bacteria.

Key words: tobacco, starch-degrading bacteria, strain screening, strain isolation, strain identification, medium optimization, fermentation conditions

中图分类号:

S572
Q939.9

周航, 孙梦瑶, 杨艾勇, 余洋洋, 裘垚, 杨永锋, 陈红丽. 一株新型烟叶淀粉降解菌的筛选鉴定及其发酵条件优化[J]. 浙江农业学报, 2026, 38(2): 206-216.

ZHOU Hang, SUN Mengyao, YANG Aiyong, YU Yangyang, QIU Yao, YANG Yongfeng, CHEN Hongli. Screening, identification and fermentation condition optimization of a novel starch-degrading bacterium isolated from tobacco leaves[J]. Acta Agriculturae Zhejiangensis, 2026, 38(2): 206-216.

图/表 9

表1 分离得到的产淀粉酶菌株的透明圈直径(D)、菌落直径(D)与淀粉酶活性(Y)

Table 1 The diameter of the transparent zone (D), colony diameter (d) and amylase activity (Y) of the isolated amylase-producing strains

菌落编号 Colony No.	d/mm	D/mm	D/d	Y/(U·mL^-1)
K1	6.0	19	3.17	68.85±1.81 a
W1	4.5	14	3.11	37.66±1.67 c
4	5.0	15	3.00	47.07±0.88 b
6	4.5	14	3.11	44.88±1.97 b

图1 菌株的生长曲线

Fig.1 Growth curve of test strain

图2 基于菌株16S RDNA基因序列构建的系统发育树

Fig.2 The constructed phylogenetic tree based on the sequences of 16S rDNA gene of the test strain

图3 碳源、氮源和无机盐种类对菌株淀粉酶活性的影响柱上无相同字母的表示差异显著(p<0.05)。下同。CMC-Na, 羧甲基纤维素钠。

Fig.3 Effects of carbon source, nitrogen source and inorganic salt species on amylase activity of test strain Bars marked without the same letters indicate significant difference at p<0.05. The same as below. CMC-Na, Sodium carboxymethyl cellulose.

图4 最适碳源、氮源和无机盐质量浓度对菌株淀粉酶活性的影响

Fig.4 Effects of mass concentration of suitable carbon source, nitrogen source and inorganic salt on amylase activity of test strain

表2 Box-Behnken试验设计与结果

Table 2 Design and results of Box-Behnken test

编号No.	A/(g·L^-1)	B/(g·L^-1)	C/(g·L^-1)	Y/(U·mL^-1)
1	5	5	13	109.238
2	15	5	13	98.694
3	5	15	13	108.919
4	15	15	13	106.970
5	5	10	9	105.924
6	15	10	9	94.843
7	5	10	17	102.993
8	15	10	17	92.286
9	10	5	9	99.207
10	10	15	9	107.173
11	10	5	17	93.949
12	10	15	17	95.265
13	10	10	13	119.450
14	10	10	13	117.574
15	10	10	13	117.348
16	10	10	13	115.780
17	10	10	13	116.785

图5 麦麸、豆粕、KH2PO4质量浓度交互作用对菌株淀粉酶活性影响的响应面和等高线图 A,麦麸质量浓度;B,豆粕质量浓度;C,KH2PO4质量浓度。

Fig.5 Response surface and contour plots of the interaction of mass concentration of wheat bran, soybean meal and KH2PO4 on amylase activity of test strain A, Mass concentration of wheat bran; B, Mass concentration of soybean meal; C, Mass concentration of KH2PO4.

图6 发酵条件对菌株淀粉酶活性的影响

Fig.6 Effect of fermentation conditions on amylase activity of test strain

表3 不同条件下烟叶的常规化学成分

Table 3 Chemical composition of tobacco leaves under different treatments

处理 Treatment	烟碱含量/ (g·kg^-1) Nicotine content/ (g·kg^-1)	还原糖含量/ (g·kg^-1) Reducing sugar content/ (g·kg^-1)	总糖含量/ (g·kg^-1) Total sugar content/ (g·kg^-1)	总氮含量/ (g·kg^-1) Total nitrogen content/ (g·kg^-1)	钾含量/ (g·kg^-1) Potassium content/ (g·kg^-1)	氯含量/ (g·kg^-1) Chlorine content/ (g·kg^-1)	淀粉含量/ (g·kg^-1) Starch content/ (g·kg^-1)	钾氯比 Ratio of potassium to chloride	两糖比 Ratio of reducing sugar to total sugar	糖碱比 Ratio of total sugar to nicotine
CK	19.5± 0.3 a	256.0± 2.0 b	296.3± 1.5 b	17.3± 0.2 b	14.3± 0.2 b	5.8± 0.1 b	61.7± 0.5 a	2.45± 0.06 b	0.86± 0.01 a	15.17± 0.24 b
T	18.1± 0.2 b	284.3± 1.5 a	323.3± 4.6 a	20.1± 0.2 a	19.0± 0.1 a	6.5± 0.2 a	31.2± 0.7 b	2.93± 0.08 a	0.88± 0.01 a	17.90± 0.32 a

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一株新型烟叶淀粉降解菌的筛选鉴定及其发酵条件优化

Screening, identification and fermentation condition optimization of a novel starch-degrading bacterium isolated from tobacco leaves

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