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

doi:10.3969/j.issn.1004-1524.20250222

Acta Agriculturae Zhejiangensis ›› 2026, Vol. 38 ›› Issue (2): 206-216.DOI: 10.3969/j.issn.1004-1524.20250222

• Crop Science • Previous Articles Next Articles

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

Abstract

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

CLC Number:

S572
Q939.9

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.

Figures/Tables 9

References 32

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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

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

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

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

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

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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

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