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Article

Antifungal Effects of Fermented Sophora flavescens and Eleutherococcus sessiliflorus Extract

by
Ju Yeon Kim
1,†,
Min Joo Chae
1,†,
Yun Gon Son
1,
Su Min Jo
1,
Na Rae Kang
1,
Seong Doo Kang
1,
Kwang Dong Kim
2,
Sang Won Lee
1,* and
Jeong Yoon Kim
1,*
1
Department of Pharmaceutical Engineering, Institute of Agricultural and Life Science (IALS), Anti-Aging Bio Cell Factory Regional Leading Research Center (ABC-RLRC), Gyeongsang National University, Jinju 52725, Republic of Korea
2
Division of Applied Life Science (BK21 Four), Anti-Aging Bio Cell Factory Regional Leading Research Center (ABC-RLRC), Gyeongsang National University, Jinju 52828, Republic of Korea
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Appl. Sci. 2024, 14(10), 4074; https://doi.org/10.3390/app14104074
Submission received: 16 April 2024 / Revised: 6 May 2024 / Accepted: 7 May 2024 / Published: 10 May 2024
(This article belongs to the Special Issue Advances in Biological Activities and Application of Plant Extracts)
Browse Figures
Versions Notes

Abstract

:
In this study, a microbial strain was isolated from humus soil to ferment Sophora flavescens and Eleutherococcus sessiliflorus extracts. The isolated microbial was identified as the Bacillus genus by 16S rRNA sequence analysis. The fermented plant extracts exhibited antifungal effects against four types plant pathogen, P. carotorum, B. cinerea, C. fructicola Sau-3, and C. gloeosporioides, according to incubation time. In particular, the fermented plant extracts showed the most activity for Colletotrichum genus in inhibiting mycelium growth. Metabolite changes in fermented S. flavescens and E. sessiliflorus extracts were confirmed through LC-Q-TOF/MS. Flavonoid and peptide derivatives were improved in fermented S. flavescens and E. sessiliflorus extracts compared to their unfermented counterparts. This study suggested that isolated Bacillus microbial fermentation could be a valuable tool in improving the bioactivity of S. flavescens and E. sessiliflorus extracts, with the potential to form more environmentally friendly antifungal agents.

1. Introduction

Plant extracts are known for promising potential antimicrobial activity against microorganisms and fungi. Many studies have reported the antimicrobial properties. One such study investigated the antimicrobial properties of water and ethanolic extracts from plants such as roselle, rosemary, clove, and thyme, against a range of foodborne pathogens and spoilage microorganisms [1,2,3]. Microbial activities occur in characterized metabolites such as terpenes, flavones, flavonols, alkaloids, and phenylpropanoids [4,5]. In recent years, there has been growing interest in improving antibacterial activity through efficient metabolite changes [6,7,8]. Several research studies have been presented, showcasing innovative methods of enhancing the antioxidant and antibacterial properties of natural substances. For instance, the fermentation of Salvia miltiorrhiza roots by Aspergillus oryzae has been shown to significantly increase its antioxidant and antibacterial activity [9]. Additionally, the development of a colorimetric sensor with high selectivity for antibacterial agents using green synthesis of gold nanocubes (utilizing sunlight and horsetail leaf extract) has shown promising results [10]. Furthermore, the antibacterial activity of olive fruit extract has been improved through the use of silver nanoparticles [11]. These advancements in antibacterial research hold great potential for the development of more effective and eco-friendly solutions.
Sophora flavescens exhibits a variety of pharmacological properties, including anti-inflammatory, antioxidant, and anticancer activities [12,13]. The root contains a range of secondary metabolites, including isoprenoid flavonoids such as kushenol, kurarinone, and maackiain, as well as alkaloids and other flavonoids [14]. The key to these beneficial effects lies in the diverse array of compounds found within S. flavescens, with flavonoids and alkaloids being the major components. Through extensive research, over 200 compounds have been isolated from the plant, and at least 50 of these have been found to possess potent antifungal properties [15]. Notably, alkaloids such as oxymatrine and matrine, extracted from S. flavescens, have been identified as having significant pharmacological effects and are commonly used in the treatment of inflammatory conditions [16].
Eleutherococcus sessiliflorus, also known as Acanthopanax sessiliflorus, is a shrub or small tree native to Eastern Asia [17]. It has been used in traditional medicine for its various properties [18]. Some of the properties associated with E. sessiliflorus include immune-stimulating, anti-inflammatory, and anticancer activities [19]. It has been found to inhibit receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclast differentiation and has been evaluated for its potential to prevent osteoporosis [20,21]. The chemical constituents of E. sessiliflorus include triterpenoids, phenolic compounds, alkaloids, flavonoids, and other bioactive compounds. These constituents have been found in various parts of the plant, including the leaves, roots, and fruits [22,23]. Some specific compounds that have been identified include chiisanoside, chiisanogenin, eleutherosides, 3,4-seco-lupanetype triterpenoids, elesesterpenes AK, and lignans [24].
Bacillus fermentation is a process in which bacteria from the Bacillus genus are used to produce a variety of products through the breakdown of organic materials. Bacillus bacteria are aerobic, Gram-positive soil bacteria that have been used for decades in the production of foods such as fermented soybean products, cheese, and yogurt [25]. During fermentation, the bacteria utilize the nutrients in the organic material to produce various substances, including enzymes, organic acids, and other metabolites. These substances can have beneficial effects on health and are used in various applications, such as food, agriculture, and industry [26]. Examples of substances produced through Bacillus fermentation include nattokinase, phytase, amylase, protease, and biosurfactants [27,28]. The process is generally carried out in liquid or solid-state fermentation systems under controlled conditions to maximize product yield and quality.
In this study, Phytophthora cactorum, Botrytis cinerea, Colletotrichum fructicola Sau-3, and Colletotrichum gloeosporioides were used to confirm the antifungal activities of bacillus-fermented S. flavescens and E. sessiliflorus extracts. P. cactorum is a well-known and significant pathogen in the field of agriculture; it is known particularly for its detrimental effects on strawberries and other fruit crops [29]. This pathogen is often transported through plant material as a latent infection and can also survive in soil and plant debris [30]. In recent years, P. cactorum has caused significant losses in the strawberry industry in Finland and has also been responsible for an increase in culling of silver birch seedlings in forest nurseries due to stem lesions [31]. The impact of this pathogen on the agricultural sector cannot be overlooked and requires immediate attention for effective management and control.
B. cinerea is a fungal pathogen that is known to cause a soft decay of plant tissues, accompanied by the growth of fuzzy gray–brown mold [32]. It can have a significant impact on many plants, particularly those grown in humid conditions. The pathogen typically enters the plant’s tissue through injuries, where it forms gray, velvety mats of sporulating tissues [33]. This can result in the death of infected twigs, causing them to die back several inches. Additionally, infected blossoms can lead to increased fruit drop and injuries to the developing fruit.
C. gloeosporioides is a well-known fungal plant pathogen that is responsible for causing bitter rot in a variety of crops worldwide, with a particular impact on perennials in tropical regions [34]. This pathogen is known to infect a range of important host plants, including citrus, yam, papaya, avocado, coffee, eggplant, sweet pepper, and tomato. In fruits, it commonly causes black or brown lesions, while in inflorescence, blight, necrosis, and lesions with flecks and streaks can occur. The symptoms produced by this fungus vary depending on the host species and the infected tissue. It is considered the most widespread and serious post-harvest disease of several tropical fruits, such as mango, papaya, pitaya, and avocado [35]. The most damaging phase of this disease is when it remains dormant and undetected, particularly during the preclimacteric phase of fruit development.
C. fructicola, commonly known as Sau-3, is a well-known and extensively studied fungus belonging to the C. gloeosporioides complex [36]. This polyphagous fungus has been reported from all five continents, causing anthracnose, bitter rot, and leaf spotting diseases on over 90 cultivated and non-cultivated woody or herbaceous plant species [37]. Its wide distribution and ability to infect a diverse range of plants make it a significant pathogen, causing significant economic losses to important crops such as apple, pear, strawberry, and other Rosaceae and non-Rosaceae species [38]. This fungus has been reported to cause anthracnose on various plants in different regions, including white jute in China [39], cassava in Brazil [40], tea in Taiwan [41], and shine muscat in Korea [42]. In southern Brazil and Uruguay, it is the primary cause of apple bitter rot and Glomerella leaf spot [43,44]. Furthermore, C. fructicola has been identified as a major pathogen of strawberries worldwide, exhibiting strong pathogenicity to both leaves and petioles [45]. Its broad host range and ability to cause severe diseases make it a significant concern for farmers and researchers alike.
This study aimed to verify the antifungal activities of S. flavescens and E. sessiliflorus through fermentation by Bacillus species. A single microbial strain was isolated from the humus soil to ferment S. flavescens and E. sessiliflorus. The fermented extract showed efficacy compared with unfermented plant extracts by exerting antifungal effects on Phytophthora cactorum, Botrytis cinerea, Colletotrichum gloeosporioides, and Colletotrichum fructicola Sau-3. The plant extracts and their fermented extracts were analyzed using LC-Q-TOF/MS to identify metabolite changes.

2. Materials and Methods

2.1. Isolation of Microbial Strains

The fermentation strain used in this study was isolated from the humus from filed soil in Gyeongsang National University (35°10′50.3″ N 128°05′38.1″ E, Jinju-si, Gyeongsang-nam-do, Republic of Korea). Sterilized water (9 mL) was added to the collected humus soil (1 g) using a sonicator. Then, 1 mL of the solution was diluted with 9 mL of sterilized water. The diluted humus solution (200 µL) was spread on starch plate medium, which was incubated for 18 h at 37 °C. The colonies observed on the plate were segregated based on their type and then cultured. After the culturing, the amylase, CMCase, and protease activities were measured based on the size of the clear zone of the separated colonies, which were sprayed with 0.05% KI-I solution. Single strains with highly effective enzymes were isolated to use for fermentation of the plant sample.

2.2. 16S rRNA Sequence Analysis

Genomic DNA of the isolated microorganisms was extracted using a genomic DNA extraction kit (Bioneer, Daejeon, Republic of Korea). The reaction mixture consisted of genomic DNA (1 µL), universal primers (27F/1492R, 5 µM), DNA polymerase, dNTPs, and PreMix (20 µL, AccuPower PCR PreMix, Bioneer), and PCR was performed. The PCR conditions were as follows; initial denaturation (95 °C, 2 min), denaturation (95 °C, 30 min), annealing (55 °C, 30 s), and extension (72 °C, 30 s) for a total of 30 cycles. After the cycles, extension was carried out at 72 °C for 5 min. An Accuprep TM PCR purification Kit (Bioneer) was used to remove and purified the remaining primers, nucleotides, polymerase, and salts. Then, 30 µL of the elution buffer (10 mM Tris-HCl, pH 8.5) was added. The base sequences of purified PCR products were analyzed by Macrogen Co., Ltd. (Seoul, Republic of Korea).

2.3. Fermentation and Cultivation of Plant Extracts

The dried roots of Sophora flavescens and Eleutherococcus sessiliflorus were purchased from a local market in Haman-gun, Gyeongsangnam-do, Republic of Korea. Each 200 g of the dried roots of S. flavescens and E. sessiliflorus was extracted with 1 L of distilled water using a sonicator for 7 h at 80 °C to make S. flavescens and E. sessiliflorus extract samples. The LB broth (0.5 g) was added to 20 mL of the plant extracts (S. flavescens and E. sessiliflorus). The pre-cultured strain was inoculated with 1 mL based on the optical density (O.D.) at 0.7. The mixture was incubated for 48 h at 37 °C. Then, it was obtained via filtration through filter paper (No. 2, Whatman, Maidstone, UK).

2.4. Antifungal Activity

The plant extract, fermented plant extract, and pre-cultured strain broth were measured for antifungal activity against Phytophthora cactorum, Botrytis cinerea, Colletotrichum gloeosporioides, and Colletotrichum fructicola Sau-3 by monitoring the inhibition zone. Phytophthora cactorum (Lebert & Cohn) Schroeter (KACC 40166), Botrytis cinerea Pers. (KACC 40573), and Colletotrichum gloeosporioides (Penzig) Penzig & Saccardo (KACC 40003) were distributed from Korean Agricultural Culture Collection (KACC, Wanju-gun, Jeollabuk-do, Republic of Korea). Colletotrichum fructicola Sau-3 was provided by Gyeongsangnam-do Agricultural Research & Extension Services at Jinju-si, Gyeongsangnam-do, Republic of Korea. Cork borer (5 mm) holes were drilled in the center and edges of the high-pressure sterilized PDA (potato dextrose agar) plate. Then, 180 μL of fungal strain was dispensed into the center hole. The plant extracts, fermented plant extracts, and pre-cultured strain medium were centrifuged at 12,000 rpm for 3 min to obtain the supernatant, which was filtered through a 0.45 μm membrane filter. Then, the filtered samples (160 µL) were loaded into three edge holes on the fungal inoculated plate. The plate was incubated for 48 h at 25 °C to observe the inhibition zone as antifungal effects of the samples.

2.5. LC-Q-TOF/MS Analysis

Identification of the metabolites from the plant extract was performed by LC-Q-TOF/MS equipped with high-performance liquid chromatography (HPLC, Shimadzu, Kyoto, Japan) and quadrupole time-of-flight mass spectrometry (Q-TOF/MS, X500R, AB Sciex, Framingham, MA, USA). The mobile phases A and B were water containing 0.1% acetic acid and acetonitrile containing 0.1% acetic acid, respectively. An Infinity Lab Poroshell 120 C18 column (2.1 × 100 mm, 2.7 μm, Agilent Technology, Santa Clara, CA, USA) was used as the analytical column. The mass ionization method used was electrospray ionization (ESI) in a positive mode, with the following source parameters: ion spray voltage, 5500 V; ion source temperature, 450 °C; curtain gas pressure, 30 psi; nebulizer gas pressure, 50 psi; heating gas pressure, 50 psi; and declustering potential, 50 V. The mass detection range was configured from m/z 50 to 1000. The resulting MS data were acquired by utilizing SCIEX OS 3.1.6 software.

2.6. Statistical Analysis

All experiments were conducted in triplicate to ensure accuracy and reproducibility. The antifungal activity data (the degree of inhibition of fungi growth) was collected using Sigma Plot (Version 10.0), which allowed for reliable statistical analysis of the results obtained from the experiments. The experimental results were then compared using a t-test, with a p-value less than 0.05 considered statistically significant enough to determine any differences between samples or treatments used in the experiment.

3. Results and Discussion

3.1. Identification of the Isolated Microbial Strains

The products amplified by 16S ribosomal RNA PCR were determined by DNA sequencing data identification of the GenBank database (http://www.ncbi.nlm.nih.gov) (accessed on 1 March 2023). As shown in Figure 1, a phylogenetic tree was observed with the Bacillus, Lactobacillus, and Streptomyces genus. The dominance of Bacillus, Lactobacillus, and Streptomyces was 93%, 98%, and 89%, respectively. Furthermore, the isolated bacteria (SM38b) was determined to be a similar strain to Bacillus amyloliquefaciens and Bacillus velezensis. The Bacillus genus is a diverse group of bacteria that play important roles in various environments. Bacillus subtilis and Bacillus cereus have been found in grass, soil, and the gastrointestinal tracts of ruminants, humans, and sponges, respectively. In this study, Bacillus amyloliquefaciens and Bacillus velezensis were isolated from humus, a nutrient-rich layer of soil. These bacteria are frequently found in soil environments and have been found to have beneficial effects on plant growth. Bacillus amyloliquefaciens, as a Gram-positive bacterium, is known for its biocontrol abilities. It has been shown to effectively combat plant root pathogens in various agricultural settings. Similarly, Bacillus velezensis has the ability to inhibit the growth of plant pathogens by competing with other bacteria. These bacteria can also produce bacterial metabolic active substances that help strengthen the plant’s immune system. Thus, Bacillus velezensis can contribute to promoting plant growth, suppressing pathogens, and maintaining the balance of the soil ecosystem.

3.2. Antifungal Effects of the Fermented and Unfermented Plant Extracts

S. flavescens and E. sessiliflorus extracts were inoculated with the isolated microbial strain to prepare the fermented plant extracts. The fermented and unfermented S. flavescens and E. sessiliflorus extracts were tested for their antifungal effects against four plant pathogens including Phytophthora cactorum, Botrytis cinerea, Colletotrichum fructicola Sau-3, and Colletotrichum gloeosporioides according to incubation times of 24, 48, 72, 96, and 120 h, respectively.
P. cactorum is a soilborne pathogen that can infect many plant species. It causes various types of damage, including leaf spots, shoot blight, root rots, and crown rots [29,46]. B. cinerea is a destructive fungal pathogen that affects numerous plant hosts. It causes fuzzy gray-brown mold and soft decay of plant tissues [47,48]. C. fructicola is known to cause anthracnose, bitter rot, and leaf spotting diseases on over 90 cultivated and non-cultivated woody or herbaceous plant species. The pathogen has been reported from all five continents and affects a wide range of crops, including apple, pear, strawberry, mango, avocado, and coffee [36,49]. C. gloeosporioides is a fungal pathogen that causes anthracnose and fruit rotting diseases in a wide range of host plants [34].
As shown in Figure 2 and Table 1, the fermented S. flavescens and E. sessiliflorus extracts showed more active antifungal activities against four fungi than the unfermented extracts. The growth inhibitory potential of P. cactorum was slightly exhibited by the effects of fermented S. flavescens extract (FSE) and fermented E. sessiliflorus extract (FEE). FSE and FEE exhibited the same inhibition patterns against P. cactorum, as follows: no inhibition at 24 h, mycelial growth inhibition with less than 2 mm at 48 h, and more than 2 mm from 96 h. Similarly, B. cinerea growth was inhibited by less than 2 mm up to 72 h and more than 2 mm thereafter by FSE and FEE treatments. The most inhibition of FSE and FEE was observed against the Colletotrichum genus (C. fructicola Sau-3 and C. gloeosporioides). For the inhibition of C. fructicola Sau-3, we observed that mycelial growth was hindered at a level of more than 2 mm by 96 h; moreover, it was inhibited by more than 3 mm when cultured for up to 120 h. The growth of C. gloeosporioides was inhibited by approximately 2 mm for up to 72 h, but FSE inhibited its growth by over 3 mm. The effects of FEE confirmed that C. gloeosporioides was suppressed at the 2 mm level throughout the entire culture period. These findings suggested that the fermented extracts of S. flavescens and E. sessiliflorus have potential as antifungal agents against these plant pathogenic fungi.

3.3. LC-Q-TOF/MS Analysis

Figure 3 shows that the peaks from the BPI gram were well separated in the unfermented and fermented S. flavescens and E. sessiliflorus extracts, according to a positive mode of LC-Q-TOF/MS. The different patterns of the peaks represented the metabolite changes of the fermented extracts compared to the control.
The BPI gram of FSE displayed (Figure 3c) predominant seven peaks (peaks 1–7), but only five peaks (peaks 1, 2, 5–7) were identified by comparing observed and theoretical mass in the individual mass gram (Table 2 and Figure 4a). Peak 1 of FSE (tR = 1.79 min) had an observed ion mass at m/z 249.1944 compared with the theoretical mass (m/z 249.1967) used to annotate matrine. Peak 2 of FSE (tR = 3.46 min) possessed a reasonable error value of −4.87 ppm, which was presented as tryptophan. Peaks 5–7 of FSE were confirmed as flavonoids, named kushenol F, G, and L, respectively. Peak 5 (tR = 14.39 min) had an error value of −4.7 ppm between the observed m/z 425.1944 and the theoretical mass at m/z 425.1964, referred to as kushenol F. Kushenol G, with the chemical formula C25H28O8, confirmed the molecular ion peaks at m/z 457.1891 [M + H]+ in the mass gram of peak 6 (tR = 15.21 min). Peak 7 (tR = 17.58 min) showed an observed mass at m/z 441.1921 with a +1.81 ppm of error value compared with the theoretical mass used to annotate kushenol L. Based on the identification of five peaks, FSE was found to contain one alkaloid, one amino acid, and three flavonoids.
FEE had six peaks (peaks 1–6) detected in the BPI gram within 30 min (Figure 3d). Among the peaks, five peaks were annotated with molecular ion peaks using individual mass grams (Table 2 and Figure 4b). Peak 1 (tR = 2.06 min) and peak 2 (tR = 3.48 min) were determined as phenylalanine and tryptophan, respectively, and the theoretical masses were found by examining the molecular ion peaks in on a mass gram. Based on the compatibility between their observed and theoretical masses, peaks 3, 4, and 6 were thought to be peptide derivatives. Peak 3 (tR = 6.24 min) displayed molecular ion peaks at m/z 262.1066 [M + H]+, indicating the presence of Asn-Glu with a theoretical mass of m/z 262.1039. Peak 4 (tR = 6.87 min) was thought to be cyclo(-Pro-Val) due to its error value of −7.10 ppm, which was determined by comparing the theoretical mass with the chemical formula (C10H16N2O2) with the observed one. Similarly, cyclo(-Phe-Pro) also displayed a molecular ion peak at m/z 245.1270 [M + H]+, which was consistent with its theoretical mass of m/z 245.1290. Thus, it was confirmed that peptide derivatives were more enhanced in FEE than in unfermented samples.

4. Conclusions

The isolated microorganism from humus soil was identified as belonging to the Bacillus genus (including species such as Bacillus amyloliquefaciens and Bacillus velezensis) due to its high dominance, which itself was based on a phylogenetic tree formed through 16S rRNA sequence analysis. The isolated Bacillus was inoculated into S. flavescens and E. sessiliflorus extracts for fermentation. The fermented plant extracts showed antifungal effects against P. carotorum, B. cinerea, C. fructicola Sau-3, and C. gloeosporioides, as plant pathogens, depending on incubation time (24, 48, 72, 96, and 120 h). The fermented plant extracts showed more active anti-fungal effects than unfermented extracts and were especially effective in inhibiting the growth of the Colletotrichum genus. Furthermore, the metabolites in the fermented plant extracts were identified by LC-Q-TOF/MS to determine the causality associated with improved biological effects. It was confirmed that the fermented S. flavescens and E. sessiliflorus extracts improved the flavonoids and peptide derivatives, respectively, based on BPI and individual mass grams. Overall, this study suggests that the fermentation-enhanced anti-fungal effects of S. flavescens and E. sessiliflorus extracts may result in the production of specific metabolites.

Author Contributions

J.Y.K. (Ju Yeon Kim) and M.J.C.: conceptualization, formal analysis, investigation; Y.G.S., S.M.J., N.R.K., S.D.K. and Y.G.S.: formal analysis, validation; K.D.K.: writing—review and editing, funding acquisition; S.W.L.: resources, writing—original draft; J.Y.K. (Jeong Yoon Kim): writing—original draft, supervision. All authors have read and agreed to the published version of the manuscript.

Funding

This work was supported by the National Research Foundation of Korea (NRF) grants funded by the Korean government (MSIT) 2021R1A5A8029490 and 2022R1F1A1063786.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The data presented in this study are available on request from the corresponding author. The data are not publicly available due to privacy and ethical reasons.

Conflicts of Interest

The authors declare no conflicts of interest.

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Applsci 14 04074 g001
Figure 1. Classification and 16S rRNA sequences of bacteria isolated from humus soil.
Figure 1. Classification and 16S rRNA sequences of bacteria isolated from humus soil.
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Figure 2. Antifungal effects of the fermented and unfermented Sophora flavescens and Eleutherococcus sessiliflorus extract against (a) Phytophthora cactorum, (b) Botrytis cinerea, (c) Colletotrichum fructicolo Sau-3, and (d) Colletotrichum gloeosporioides.
Figure 2. Antifungal effects of the fermented and unfermented Sophora flavescens and Eleutherococcus sessiliflorus extract against (a) Phytophthora cactorum, (b) Botrytis cinerea, (c) Colletotrichum fructicolo Sau-3, and (d) Colletotrichum gloeosporioides.
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Figure 3. LC-Q-TOF/MS analysis of the fermented and unfermented Sophora flavescens and Eleutherococcus sessiliflorus extract. Base peak intensity (BPI) of (a) unfermented Sophora flavescens extract, (b) fermented Sophora flavescent extracts, (c) unfermented Eleutherococcus sessiliflorus extract, and (d) fermented Eleutherococcus sessiliflorus extract.
Figure 3. LC-Q-TOF/MS analysis of the fermented and unfermented Sophora flavescens and Eleutherococcus sessiliflorus extract. Base peak intensity (BPI) of (a) unfermented Sophora flavescens extract, (b) fermented Sophora flavescent extracts, (c) unfermented Eleutherococcus sessiliflorus extract, and (d) fermented Eleutherococcus sessiliflorus extract.
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Figure 4. Mass gram of individual peaks from (a) fermented S. flavescens extract (FSE) and (b) fermented E. sessiliflorus extract (FEE) from BPI chromatograms.
Figure 4. Mass gram of individual peaks from (a) fermented S. flavescens extract (FSE) and (b) fermented E. sessiliflorus extract (FEE) from BPI chromatograms.
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Table 1. Inhibitory effects of targeted fungi by the fermented and unfermented Sophora flavescens and Eleutherococcus sessiliflorus extract.
Table 1. Inhibitory effects of targeted fungi by the fermented and unfermented Sophora flavescens and Eleutherococcus sessiliflorus extract.
Target FungiIncubation Time (h)Antifungal Effects
SE aFSE bEE cFEE d
P. cactorum24- e---
B. cinerea-+-+
C. fructicola Sau-3-++-++
C. gloeoporioides-++-++
P. cactorum48++-+
B. cinerea-+-+
C. fructicola Sau-3-++-++
C. gloeoporioides-++-++
P. cactorum72++-+
B. cinerea+++-++
C. fructicola Sau-3-++-++
C. gloeoporioides-++-++
P. cactorum96+++-++
B. cinerea+++-++
C. fructicola Sau-3-++-++
C. gloeoporioides-+++-++
P. cactorum120+++-++
B. cinerea+++-++
C. fructicola Sau-3-+++-+++
C. gloeoporioides-+++-++
a SE means Sophora flavescens extract, b FSE means fermented Sophora flavescens extract, c EE means Eleutherococcus sessiliflorus extract, and d FEE means fermented Eleutherococcus sessiliflorus extract. e The effectiveness of antifungal activities is expressed depending on the degree of inhibition of fungi growth. The inhibition is displayed as +, ++, and +++, referring to less than 2 mm, less than 3 mm, and more than 3 mm in comparison to no inhibition, respectively.
Table 2. Identification of metabolites from the fermented plant extracts using LC-TOF/MS.
Table 2. Identification of metabolites from the fermented plant extracts using LC-TOF/MS.
No.tR
(min)		Observed
Ion (m/z)		Theoretical
Ion (m/z)		Error
(ppm)		AdductsFormulaIdentification
Fermented Sophora flavescens Extract
11.79249.1944249.1967−9.23M + HC15H24N2OMatrine
23.46205.0967205.0977−4.87M + HC11H12N2O2Tryptophan
311.72553.3334----Unknown
412.72560.2692----Unknown
514.39425.1944425.1964−4.70M + HC25H28O6Kushenol F
615.21457.1891457.1862+6.34M + HC25H28O8Kushenol G
717.58441.1921441.1913+1.81M + HC25H28O7Kushenol L
Fermented Eleutherococcus sessiliflorus extract
12.06166.0851166.0868−10.24M + HC9H11NO2Phenylalanine
23.48205.0965205.0977−5.85M + HC11H12N2O2Tryptophan
36.24262.1066262.1039+10.30M + HC9H15N3O6Asn-Glu
46.87197.1276197.1290−7.10M + HC10H16N2O2Cyclo(-Pro-Val)
512.87561.2748----Unknown
614.31245.1270245.1290−8.16M + HC14H16N2O2Cyclo(-Phe-Pro)
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Kim, J.Y.; Chae, M.J.; Son, Y.G.; Jo, S.M.; Kang, N.R.; Kang, S.D.; Kim, K.D.; Lee, S.W.; Kim, J.Y. Antifungal Effects of Fermented Sophora flavescens and Eleutherococcus sessiliflorus Extract. Appl. Sci. 2024, 14, 4074. https://doi.org/10.3390/app14104074

AMA Style

Kim JY, Chae MJ, Son YG, Jo SM, Kang NR, Kang SD, Kim KD, Lee SW, Kim JY. Antifungal Effects of Fermented Sophora flavescens and Eleutherococcus sessiliflorus Extract. Applied Sciences. 2024; 14(10):4074. https://doi.org/10.3390/app14104074

Chicago/Turabian Style

Kim, Ju Yeon, Min Joo Chae, Yun Gon Son, Su Min Jo, Na Rae Kang, Seong Doo Kang, Kwang Dong Kim, Sang Won Lee, and Jeong Yoon Kim. 2024. "Antifungal Effects of Fermented Sophora flavescens and Eleutherococcus sessiliflorus Extract" Applied Sciences 14, no. 10: 4074. https://doi.org/10.3390/app14104074

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