Isolation and Identification of a High-yielding Magnolol and Honokiol Strain from Magnolia officinalis and Optimization of the “Sweating” Process

doi:10.13560/j.cnki.biotech.bull.1985.2025-0422

Abstract

Abstract:

Objective To clarify the correlation between the microbial colonies and the quality of Magnolia officinalis during the process of “sweating”, and to investigate the regulatory mechanisms of key microbial species on the biosynthesis and accumulation of magnolol and honokiol in M. officinalis. Method Traditional culture separation and HPLC were systematically employed to isolate and screen functionally critical microbial strains during the “sweating” process of M. officinalis. The target strains were identified using molecular biological techniques, and phylogenetic trees were constructed to analyze their evolutionary status and genetic relationships. The process parameters of “sweating” of strain HP3 were optimized by using single-factor experiments combined with the Box-Behnken response surface method. Result Forty-eight strains were isolated and purified from the “sweating” M. officinalis samples. Subsequent screening revealed 9 strains that significantly increased the production of magnolol and honokiol. Among these, strain HP3 presented the most pronounced efficacy and was identified as Bacillus velezensis. The optimization of the “sweating” process parameters for strain HP3 revealed the following optimal conditions: Sweating temperature 31.84 ℃, sweating time 1.9 d, boiling time 6.37 min and drying temperature 80 ℃. Under these conditions, the total amount of magnolol and honokiol was as high as 5.149%, an increase of 62.67% compared with the non-sweated samples. Conclusion A strain HP3 was isolated and identified from the “sweating” process of M. officinalis, which demonstrated remarkable capability in enhancing the total content of magnolol and honokiol. Furthermore, the optimal sweating process parameters for strain HP3 were determined through response surface optimization experiments. The study has revealed the bacterial communities closely related to the quality of M. officinalis during its “sweating” process, providing an important basis for the development of microbial resources of M. officinalis medicinal materials.

Key words: Magnolia officinalis, isolation and identification, magnolol, honokiol, “sweating”

ZHANG Ru, LI Yi-ming, ZHANG Tong-xi, SUN Zhan-bin, REN Qing, PAN Han-xu. Isolation and Identification of a High-yielding Magnolol and Honokiol Strain from Magnolia officinalis and Optimization of the “Sweating” Process[J]. Biotechnology Bulletin, 2025, 41(8): 322-334.

Figures/Tables 15

References 35

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属 Genus	筛选培养基 Screening medium	稀释梯度 Dilution gradient	数量 Number
Bordetella（鲍特氏菌属）	TSA	10^-3	1
Alcaligenes（产碱杆菌属）	TSA/BHI	10^-4/10^-5	4
Brevundimonas（短波单胞菌属）	TSA/BHI	10^-4/10^-5	3
Brucella（布鲁氏菌属）	TSA	10^-4	1
Stenotrophomonas（黄单胞菌属）	NA/TSA/BHI/R₂A	10^-4/10^-5	5
Acinetobacter（不动杆菌属）	NA/TSA/BHI/R₂A	10^-4/10^-5	4
Pseudomonas（假单胞菌属）	NA/BHI	10^-4	3
Enterobacter（肠杆菌属）	R₂A	10^-4	1
Klebsiella（克雷伯氏菌属）	R₂A	10^-4/10^-5/10^-6	4
Leclercia（勒克氏菌属）	NA	10^-4	1
Curtobacterium（短小杆菌属）	NA/TSA	10^-4/10^-5	3
Georgenia（乔治菌属）	NA/TSA/BHI	10^-3/10^-4/10^-5	5
Brevibacterium（短杆菌属）	NA/TSA/BHI	10^-3/10^-4/10^-5	4
Streptomyces（链霉菌属）	TSA	10^-4	1
Chryseobacterium（金黄杆菌属）	NA	10^-4	1
Staphylococcus（葡萄球菌属）	NA	10^-4	1
Lysinibacillus（赖氨酸芽胞杆菌属）	NA/TSA	10^-4	2
Sporosarcina（八叠球菌属）	TSA	10^-4	1
Oceanobacillus（大洋芽胞杆菌属）	TSA	10^-5	1
Bacillus（芽胞杆菌属）	TSA/BHI	10^-3	2

属 Genus	筛选培养基 Screening medium	稀释梯度 Dilution gradient	数量 Number
Bordetella（鲍特氏菌属）	TSA	10^-3	1
Alcaligenes（产碱杆菌属）	TSA/BHI	10^-4/10^-5	4
Brevundimonas（短波单胞菌属）	TSA/BHI	10^-4/10^-5	3
Brucella（布鲁氏菌属）	TSA	10^-4	1
Stenotrophomonas（黄单胞菌属）	NA/TSA/BHI/R₂A	10^-4/10^-5	5
Acinetobacter（不动杆菌属）	NA/TSA/BHI/R₂A	10^-4/10^-5	4
Pseudomonas（假单胞菌属）	NA/BHI	10^-4	3
Enterobacter（肠杆菌属）	R₂A	10^-4	1
Klebsiella（克雷伯氏菌属）	R₂A	10^-4/10^-5/10^-6	4
Leclercia（勒克氏菌属）	NA	10^-4	1
Curtobacterium（短小杆菌属）	NA/TSA	10^-4/10^-5	3
Georgenia（乔治菌属）	NA/TSA/BHI	10^-3/10^-4/10^-5	5
Brevibacterium（短杆菌属）	NA/TSA/BHI	10^-3/10^-4/10^-5	4
Streptomyces（链霉菌属）	TSA	10^-4	1
Chryseobacterium（金黄杆菌属）	NA	10^-4	1
Staphylococcus（葡萄球菌属）	NA	10^-4	1
Lysinibacillus（赖氨酸芽胞杆菌属）	NA/TSA	10^-4	2
Sporosarcina（八叠球菌属）	TSA	10^-4	1
Oceanobacillus（大洋芽胞杆菌属）	TSA	10^-5	1
Bacillus（芽胞杆菌属）	TSA/BHI	10^-3	2

来源 Source	离差平方和 Sum of squared	自由度 Freedom	均方 Mean	F值 F value	P值 P value	显著性 Significance
模型 Model	9.55	14	0.682 4	5.39	0.002 9	Significant
A	0.655 5	1	0.655 5	5.17	0.042 1	Significant
B	0.922 5	1	0.922 5	7.28	0.019 4	Significant
C	1.16	1	1.160	9.12	0.010 7	Significant
D	0.307 2	1	0.307 2	2.42	0.145 5
AB	0.032 1	1	0.032 1	0.253 6	0.623 7
AC	0.002 5	1	0.002 5	0.019 7	0.890 6
AD	0.072 1	1	0.072 1	0.568 7	0.465 3
BC	0.126 9	1	0.126 9	1.00	0.336 7
BD	0.359 4	1	0.359 4	2.84	0.118 0
CD	0.828 3	1	0.828 3	6.54	0.025 2	Significant
A²	3.19	1	3.190	25.13	0.000 3	Significant
B²	3.51	1	3.510	27.67	0.000 2	Significant
C²	0.899 5	1	0.899 5	7.10	0.020 6	Significant
D²	0.312 9	1	0.312 9	2.47	0.142 1
残差 Residual	1.52	12	0.126 7
失拟项 Lack of fit	1.17	10	0.117 1	0.670 2	0.729 0	Non-significant
纯误差 Pure error	0.349 5	2	0.174 8
总和 Total sum	11.07	26
R²=0.862 7

来源 Source	离差平方和 Sum of squared	自由度 Freedom	均方 Mean	F值 F value	P值 P value	显著性 Significance
模型 Model	9.55	14	0.682 4	5.39	0.002 9	Significant
A	0.655 5	1	0.655 5	5.17	0.042 1	Significant
B	0.922 5	1	0.922 5	7.28	0.019 4	Significant
C	1.16	1	1.160	9.12	0.010 7	Significant
D	0.307 2	1	0.307 2	2.42	0.145 5
AB	0.032 1	1	0.032 1	0.253 6	0.623 7
AC	0.002 5	1	0.002 5	0.019 7	0.890 6
AD	0.072 1	1	0.072 1	0.568 7	0.465 3
BC	0.126 9	1	0.126 9	1.00	0.336 7
BD	0.359 4	1	0.359 4	2.84	0.118 0
CD	0.828 3	1	0.828 3	6.54	0.025 2	Significant
A²	3.19	1	3.190	25.13	0.000 3	Significant
B²	3.51	1	3.510	27.67	0.000 2	Significant
C²	0.899 5	1	0.899 5	7.10	0.020 6	Significant
D²	0.312 9	1	0.312 9	2.47	0.142 1
残差 Residual	1.52	12	0.126 7
失拟项 Lack of fit	1.17	10	0.117 1	0.670 2	0.729 0	Non-significant
纯误差 Pure error	0.349 5	2	0.174 8
总和 Total sum	11.07	26
R²=0.862 7