竹屑制备天然防腐剂的工艺优化及抑菌性能研究

胡永乐; 张传海; 张媛; 尤永芳; 黄薇; 贾小丽

doi:10.19303/j.issn.1008-0384.2020.02.014

竹屑制备天然防腐剂的工艺优化及抑菌性能研究

doi: 10.19303/j.issn.1008-0384.2020.02.014

武夷学院生态与资源工程学院/福建省生态产业绿色技术重点实验室，福建　武夷山　354300

基金项目: 福建省自然科学基金项目（2019J01827）；福建省生态产业绿色技术重点实验室开放课题基金项目（WYKF2018-6）；国家级大学生创新创业训练计划项目（201810397005）；南平市科技计划项目（N2017DN07）

详细信息

作者简介:
胡永乐（1986−），女，硕士，讲师，研究方向：天然产物开发和利用（E-mail：huyongle114@163.com）

通讯作者:
贾小丽（1980−），女，博士，教授，研究方向：植物生理与分子生态学研究（E-mail：jcjyx@126.com）

中图分类号: TS202.3
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- 被引次数: 0
出版历程
- 收稿日期: 2019-08-02
- 修回日期: 2019-11-01
- 刊出日期: 2020-02-01

Process Optimization and Antimicrobial Activity of Natural Preservative Extracted from Bamboo Sawdust

Fujian Provincial Key Laboratory of Eco-Industrial Green Technology, College of Ecology and Resource Engineering, Wuyi University, Wuyishan, Fujian　354300, China

摘要

摘要: 目的为研究竹屑制备天然防腐剂的最优工艺并探讨其抑菌性能。方法以竹屑为原料，采用乙酸乙酯回流法，在单因素试验基础上，应用响应面法优化天然防腐剂制备条件，并考察其抑菌活性。结果竹屑制备天然防腐剂的最佳工艺条件为超声功率300 W、提取温度89℃、液料比20 ∶ 1、提取时间2 h。在此条件下，所得提取物对大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌和黑曲霉的抑菌效果达到最强，抑菌圈直径分别为：9.33、15.67、10.00、6.67 mm，最低抑菌浓度分别为：0.025 00、0.012 50、0.050 00和0.100 00 g·mL⁻¹，竹屑天然防腐剂得率为4.77%。结论响应面法优化工艺制备的天然防腐剂对4种供试菌抑菌效果良好，制备工艺可行性高。该工艺可为竹废弃物的加工再利用提供参考。
- 竹屑 /
- 天然防腐剂 /
- 响应面分析法 /
- 工艺优化 /
- 抑菌
Abstract: Objective Optimum preparation and antibacterial properties of a natural preservative extracted from bamboo sawdust were studied. Method Utilizing bamboo sawdust, a natural preservative was prepared with the ethyl acetate reflux method. Processing conditions were optimized by the response surface methodology based on the results of a single factor test. In vitro antimicrobial activity of the extract was determined. Result The finalized process applied 300 W ultrasonic power on the mixture of solvent-to-substrate ratio at 20 ∶ 1 to extract at 89℃ for 2 h. The product yield reached 4.77%. The extracted preservative displayed peak antimicrobial effects on Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Aspergillus niger with the average inhibition diameters of 9.33 mm at a concentration of 0.025 00 g·mL⁻¹, 15.67 mm at a concentration of 0.012 5 0g·mL⁻¹, 10.00 mm at a concentration of 0.050 00 g·mL⁻¹, and 6.67 mm at a concentration of 0.100 00 g·mL⁻¹, respectively. Conclusion The natural preservative prepared by the newly developed extraction method showed strong in vitro antimicrobial effect, and the optimized process could be upscaled to fully utilize the bamboo sawdust.
- Bamboo sawdust /
- natural preservative /
- response surface methodology /
- process optimization /
- antimicrobial property

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图 1 超声功率、液料比、提取时间、提取温度对抑菌圈直径的影响

Figure 1. Effects of extraction ultrasonic power, solvent-to-substrate ratio, time, and temperature on microbial inhibition of preservative

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图 2 超声功率、液料比、提取时间、提取温度对竹屑天然防腐剂得率的影响

Figure 2. Effects of extraction ultrasonic power, solvent-to-substrate ratio, time, and temperature on yield of preservative

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图 3 提取温度与液料比的响应面和等高线分析

Figure 3. Response surface diagram and contour lines between solvent-to-substrate ratio and temperature applied for extraction

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图 4 提取温度与提取时间的响应面和等高线分析

Figure 4. Response surface diagram and contour lines between temperature and time applied for extraction

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图 5 液料比与提取时间的响应面和等高线分析

Figure 5. Response surface diagram and contour lines between solvent-to-substrate ratio and time applied for extraction

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表 1 Box-Behnken试验设计的因素及水平

Table 1. Factors and levels of Box-Behnken experimental design

水平 Level	提取温度 Extraction temperature/℃	液料比 Solvent-to-substrate ratio/（mL:g）	提取时间 Extraction time/h
−1	80	15∶1	1
0	90	20∶1	2
1	100	25∶1	3

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表 2 响应面试验设计组合及结果

Table 2. Design combinations and results of response surface test

试验号 Test number	A提取温度 Extraction temperature/℃	B液料比 Solvent-to-substrate ratio/（mL:g）	C提取时间 Extraction time/h	抑菌圈直径 Inhibitory zone diameter/mm
试验号 Test number	A提取温度 Extraction temperature/℃	B液料比 Solvent-to-substrate ratio/（mL:g）	C提取时间 Extraction time/h	大肠杆菌 E. coli	金黄色葡萄球菌 S. aureus	枯草芽孢杆菌 B. subtilis	黑曲霉 A. niger	平均值 Average
1	80	15∶1	2	3.75	6.79	2.50	3.21	4.06
2	100	15∶1	2	2.82	3.42	6.59	4.21	4.26
3	80	25∶1	2	4.24	7.41	5.38	3.58	5.15
4	100	25∶1	2	3.22	2.80	4.80	2.19	3.25
5	80	20∶1	1	3.50	8.56	1.88	2.70	4.16
6	100	20∶1	1	1.20	2.02	2.75	1.26	1.81
7	80	20∶1	3	4.58	8.96	3.02	3.76	5.08
8	100	20∶1	3	3.82	4.41	5.56	3.41	4.30
9	90	15∶1	1	2.52	3.86	4.00	2.42	3.20
10	90	25∶1	1	3.15	4.02	4.22	1.78	3.29
11	90	15∶1	3	5.49	6.54	6.78	3.92	5.68
12	90	25∶1	3	5.86	7.64	8.42	3.60	6.38
13	90	20∶1	2	8.75	16.02	10.51	7.25	10.63
14	90	20∶1	2	9.75	14.03	11.00	7.47	10.56
15	90	20∶1	2	7.75	14.98	10.03	7.01	9.94
16	90	20∶1	2	9.25	15.54	10.35	7.34	10.62
17	90	20∶1	2	9.15	16.40	9.90	7.21	10.67

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表 3 响应面方差分析

Table 3. Variance analysis of response surfaces

差异来源 Source	平方和 Sun of quares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型 Model	154.75	9	17.19	94.18	＜0.000 1**
A-提取温度Extraction temperature	2.92	1	2.92	15.97	0.005 2**
B-液料比Solvent-to-substrate ratio	0.095	1	0.095	0.52	0.494 9
C-提取时间Extraction time	10.08	1	10.08	55.21	0.000 1**
AB	1.1	1	1.1	6.04	0.043 6*
AC	0.62	1	0.62	3.38	0.108 8
BC	0.093	1	0.093	0.51	0.498 4
A²	53.12	1	53.12	290.98	＜0.000 1**
B²	31.89	1	31.89	174.67	＜0.000 1**
C²	40.32	1	40.32	220.85	＜0.000 1**
残差 Residual	1.28	7	0.18
失拟项 Lack of fit	0.9	3	0.3	3.2	0.145 5
纯误差 Pure error	0.38	4	0.094
总和 Total	156.03	16
注：表示显著差异，P＜0.05；表示极显著差异，P＜0.01。 Note: P＜0.05, the difference is significant at 0.05 level; ** P＜0.01, the difference is significant at 0.01 level.

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表 4 最低抑菌浓度的测定

Table 4. Determination of minimal inhibitory concentration for peak antimicrobial effect

抑菌浓度 Concentration /g·mL⁻¹	抑菌圈直径 Inhibitory zone diameter/mm
抑菌浓度 Concentration /g·mL⁻¹	大肠杆菌 E. coli	金黄色葡萄球菌 S. aureus	枯草芽孢杆菌 B. subtilis	黑曲霉 A. niger
0.200 00	5.24±0.83	8.06±0.62	7.64±0.85	4.06±0.72
0.100 00	4.38±0.77	6.42±0.81	5.32±0.76	1.04±0.18
0.050 00	2.18±0.22	5.02±0.57	2.14±0.35	0.00
0.025 00	1.15±0.14	3.02±0.42	0.00	0.00
0.012 50	0.00	1.04±0.15	0.00	0.00
0.006 25	0.00	0.00	0.00	0.00
空白对照 Blank control	0.00	0.00	0.00	0.00

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