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Article

Lovastatin Production by Wild Eurotium cristatum Isolated from Fuzhuan Brick Tea Produced Using Forest Resources in Auhua

by
Taotao Li
1,2,3,
Zhanjun Liu
3,
Jun Li
2,3,
Yajun Zheng
2,3,
Zhonghua Liu
1,* and
Peixue Ling
2,4,*
1
Key Lab of Education Ministry for Tea Science, National Research Center of Engineering Technology for Utilization of Botanical Functional Ingredients, Hunan Agricultural University, Changsha 410128, China
2
Hunan Weichu Furuida Biotechnology Limited Liability Company, Yiyang 413046, China
3
Hunan Provincial Key Lab of Dark Tea and Jin-Hua, Hunan City University, Yiyang 413000, China
4
National Glyceon Egineering Research Center, Shandong University, Jinan 250199, China
*
Authors to whom correspondence should be addressed.
Forests 2023, 14(7), 1409; https://doi.org/10.3390/f14071409
Submission received: 19 May 2023 / Revised: 30 June 2023 / Accepted: 6 July 2023 / Published: 11 July 2023
(This article belongs to the Special Issue Advances in the Sustainable Development and High-Value Utilization of Forestry Resources)
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Abstract

:
Fuzhuan brick tea is a dark tea produced using tea plant leaves, and its quality is strongly affected by Eurotium cristatum. The superior Eurotium cristatum strain could enhance functional components in Fuzhuan brick tea, improving its quality and economic value. To research the lipid-lowering substance of Eurotium cristatum in Fuzhuan brick tea from Anhua country tea production area, several wild-type Eurotium cristatum strains were screened for lovastatin production. Of all 20 wild-type strains, 6 Eurotium cristatum strains could produce lovastatin, and the H20 test strain could produce the highest lovastatin. The organ of lovastatin production was studied, and its yield in sexual propagule (ascospore and ascocarp) was the highest. Some fermentation parameters for lovastatin produced by the H20 strain were researched, and the optimum condition, such as PDA medium containing additional 8% soluble starch, 32 °C, and 120 r/min, are reported. These results are promising for developing one Fuzhuan brick tea with a high lipid-lowering function and expanding the scope of tea plant forest resource application.

1. Introduction

Lovastatin effectively reduces human cholesterol levels and has been widely used as a lipid-lowering drug [1,2]. It could limit cholesterol synthesis by inhibiting the 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase [3,4,5]. Lovastatin, as a secondary metabolite, is produced by numerous fungal species [6,7,8]. Lovastatin is always isolated from Monascus and Aspergillus terreus because they can produce more lovastatin. Except for Monascus and Aspergillus terreus, Eurotium cristatum can also produce lovastatin.
Eurotium cristatum is the dominant fungus involved in the fermentation of Fuzhuan brick tea, which functions to lower blood lipid levels [9,10,11,12]. Based on its secreted amylase and oxidase (such as polyphenol oxidase, cytochrome oxidase), tea leaves could transform into many compounds that benefit human health, such as anti-obesity activity, anti-diabetic activity, gastrointestinal protection, and so on. Eurotium cristatum also can secrete a lot of natural pigments which improve the sensory properties and market competitiveness of Fuzhuan brick tea. The characteristic color, fragrance, and taste of Fuzhuan brick tea are mainly derived from the transformation of tea leaves substance by Eurotium cristatum. Hence, Eurotium cristatum is an important indicator to evaluate the quality of Fuzhuan brick tea. Fuzhuan brick tea and Eurotium cristatum water extract decreased lipid droplet size and fat accumulation in Caenorhabditis elegans, zebrafish model, or high-fat diet-treated mice [13,14,15]. The safety of dark tea could also be studied by animal model experiments. Evidence from those studies indicated that dark tea can benefit human health [15]. The anti-obesity active metabolite is mostly polysaccharides, polyphenols, and small molecule compounds, including lovastatin [16,17,18]. In recent years, research has shown that Eurotium cristatum can produce lovastatin with a limited yield [19,20,21]. Based on the colonial morphology and ascocarp type, Eurotium cristatum has various strains. The lovastatin produced by various Eurotium cristatum strains currently lacks sufficient research.
Fuzhuan brick tea is a post-fermented tea produced using tea plant leaves [22,23]. The tea plant is an important economic forest [24,25,26], and it is beneficial to improve the economic value of forestry resources through manufacturing tea products. It is difficult to improve the economic value of tea plant forests by relying on traditional dark tea products. One possible way is to develop new products with higher lipid-lowering effects. Researches show that lipid-lowering effect of Anhua dark tea is related to many compounds, such as tea polysaccharide, tea polyphenols, tea amino acids, and some active small molecule substances, such as lovastatin (always lower than 20 μg/g) [27]. Compared with other dark tea, the active small molecule and microorganisms in Fuzhuan brick tea are special. Microorganisms, especially Eurotium cristatum, influence the quality of Fuzhuan brick tea. Hence, increasing the active small molecule yield in Eurotium cristatum could enhance the lipid-lowering effect of Fuzhuan brick tea. The main fungus in this tea and its effect on tea quality can provide insights into screening high-quality tea varieties. They could lay the foundation for the conservation of germplasm resources in tea trees [28,29].
Anhua County is a county-level city in Yiyang City, Hunan Province. It is located in the mid-north part of Hunan Province and in the middle reaches of the Zijiang River. Anhua has a total area of 4950 square kilometers, and its forest coverage rate is 76.51%. Anhua County has a subtropical monsoonal humid climate, with an average annual temperature and precipitation of 16.2 °C and 1622 mm, respectively. The parent material of soil in Anhua is mainly slate-weathered material and contains small sandy conglomerate, limestone, and granite-weathered material. The soil type is moderately sticky and sandy. The weak acid and rich nutrient content of soil provide sufficient conditions for the growth of tea plant forests. Many famous tea production areas include Yuntai Mountain, Furong Mountain, Gaojia Mountain, Gaojiaxi and Majiaxi Villages, Wulong Mountain, and so on. The main tea products in Anhua are dark tea. Traditional Anhua dark tea includes compressed tea (Fuzhuan brick tea, Flower-brick tea, and black-brick tea), Qianliang dark tea, and Tianjian dark tea. All types of dark tea have a certain lipid-lowering effect, and the lipid-lowering effect of Fuzhuan brick tea is best.
The tea plant is one of the important economic forests in China. The tea plant forest contains tea gardens or “wild tea” trees grove, and its acreage is over 10 km2. Developing the economic value of the Anhua tea plant forest is mainly related to new dark tea products with better sensory properties or health benefits. The present study was conducted to produce lovastatin using Aspergillus terreus or Monascus rubber. Some researchers produced the lovastatin by Eurotium cristatum, and they mainly focused on detecting its yields in the fermentation broth or dark tea. In this research, we focused on screening Eurotium cristatum with higher lovastatin yields, from Fuzhuan brick tea produced by Anhua dark tea production areas. Comprehensive studies of lovastatin production by wild Eurotium cristatum are lacking. To better understand lovastatin production in Eurotium cristatum, several wild strains with different colonial morphology were screened from Fuzhuan brick tea produced by the main dark tea production areas in Anhua country. Furthermore, the organs (mycelium, ascocarp, ascospore, conidium, and fermentation broth) of lovastatin production by Eurotium cristatum were investigated. The influences of fermentation parameters on lovastatin production also were studied to obtain suitable conditions. In the future, the screened Eurotium cristatum strains could produce more natural lovastatin in the fermentation of Fuzhuan brick tea, and one new Fuzhuan brick tea product with a greater lipid-lowering effect could be developed. These new dark tea products may enhance the utilization rate and improve the economic value of tea plant forest resources in Anhua country.

2. Materials and Methods

2.1. Anhua Tea Plant Forests Resource Site

Anhua County (110°–112° E, 27°–29° N) is a county-level city in Hunan Yiyang. The resources of tea plant forests are rich. All Fuzhuan brick tea was obtained from a special tea production mountain area in Anhua County. The first one is Yuntai Mountain (111°02′ E, 28°37′ N), and its representative tea variety has big leaves. The second is Furong Mountain (111°45′ E, 28°10′ N), whose maximum elevation is 1.4 km. The third is Gaojia Mountain (111°45′ E, 28°10′ N), whose maximum elevation is 1.2 km. The fourth is Gaojiaxi and Majiaxi villages, and their maximum elevation is 1.3 km. The fifth is the Wulong Mountain area, and its maximum elevation is 0.8 km. Except for the tea production area, tea tree variety, planting patterns, and tea processing technology also were considered to select Fuzhuan brick tea.

2.2. Strain Isolation and Sub-Culturing

Wild Eurotium cristatum was separated from obtained Fuzhuan brick tea by dilution plate method. 1 g Fuzhuan brick tea was added to 100 mL sterile water and placed in the oscillating incubator (100 r/min) for 30 min. The mixed liquor-suspended tea was diluted at a gradient of 10 times. 0.1 mL diluted tea solution was spread to potato dextrose agar (PDA) and grown at 28 °C for seven days until complete single colony formation. The grown Eurotium cristatum was further dilution cultured to purify the strain twice. The purified Eurotium cristatum strains were cultured onto slants at 28 °C for 7 days. The screened Eurotium cristatum strains were labeled based on their special colonial morphology. The slants were maintained at 4 °C and were sub-cultured periodically for further use.

2.3. Lovastatin Production by Liquid Fermentation

For fermentations, all screened Eurotium cristatum strains were first cultured at 28 °C in a seed medium (PDA medium containing additional 10% sucrose) on an incubator under the condition of a rotation rate of 120 r/min. After incubation for 5 days, 2 mL of the seed solution was added to 100 mL fermentation medium (PDA medium containing additional 4% sucrose). The temperature and rotation speeds were 32 °C and 120 r/min, respectively. Lovastatin production was detected after 8 days. All experiments were repeated three times.

2.4. Production of Ascocarp, Ascospore, and Conidium

The ascocarp and ascospore were separated from loose dark tea fermented by Eurotium cristatum H20 strain. The loose dark tea was produced by a reported method. The tea sample (20 g) was sterilized at 121 °C for 20 min and inoculated with 2 mL fungal suspension (107 CFU/mL). The Eurotium cristatum H20 strain fermented the tea at 28 °C for 10 days. The humidity during fermentation was maintained at 80%. The fermented loose dark tea was stored in a ventilated and dry environment. The conidium was formed in hyperosmotic solution. 2 mL of the seed solution was added to 100 mL PDA medium containing 15% sucrose and 10% sodium chloride, then stationary culture at 28 °C for 8 days. After fermentation, the conidium was separated from the mycoderm by degreased gauze and dried in a baking oven.

2.5. Extraction of Lovastatin

The fermented broth was centrifuged at 3000 r/min, and after discarding supernate, the mycelium (1 g) was added to 10 mL of 95% ethanol and extracted at room temperature for 2 h. In addition, 5 mL liquid supernatant was mixed with 5 mL of 95% ethanol and extracted at room temperature for 2 h. 1 g ascocarp was suspended in 5 mL water and broken by ultrasonic method at 600 W for 10 min. The mixed solution was centrifuged at 3000 r/min, and the liquid supernatant was extracted with 95% ethanol (5 mL) at room temperature for 2 h. The sediment was resuspended in 5 mL water and filtered by 4 layers of gauze. The filtrate was dried in a baking oven. And the ascospore was obtained. Ascospore or conidium was suspended in 5 mL water, diluted to 108/mL, and then broken by the glass bead-beating method [30]. After the broken wall, the mixture was extracted with 5 mL of 95% ethanol for 2 h at room temperature. All experiments were repeated three times.

2.6. Determination of Lovastatin

The lovastatin was detected by the reported HPLC method [8]. The lovastatin extracts were filtered through a 0.2 μm filter membrane and then measured by an HPLC system. The analytical spectrophotometric detector was set at 238 nm. The type of column is the EC-C18 chromatography column (2.1 × 50 mm, impacted PEEK-lined stainless steel columns), which was purchased from Agilent Technologies Co., Ltd. (Santa Clara, CA, USA). An isocratic mobile phase of methanol:water at the ratio of 85:15 (v/v) was used. The flow rate was 0.8 mL/min at 25 °C. Compared with strand lovastatin, the error of retention time of lovastatin in fermentation broth should be lower than 5%. All samples were detected three times, and the average concentration of lovastatin was calculated.

3. Results and Discussion

3.1. Lovastatin Production by Different Eurotium cristatum Strains

In the first part of the research, lovastatin production screening was carried out using different wild Eurotium cristatum strains. These strains were isolated from different Fuzhuan brick tea purchased from various Fuzhuan brick dark tea production areas in Anhua County. The isolated wild Eurotium cristatum strains were labeled based on their special colony morphologies. Among them, 20 Eurotium cristatum strains with larger different colony morphologies (length of mycelium, smoothness, and wrinkle) were selected to study the production capacity of lovastatin. Lovastatin was evaluated in 8-day-old PDA fermentation broth to screen the lovastatin production of the 20 wild tested strains. To ensure lovastatin yields in Eurotium cristatum strains, an HPLC assay was employed. The lovastatin production of different tested strains had significant variation. Among 20 Eurotium cristatum strains, only six tested strains produced lovastatin in yields ranging from 1 to 10 μg/mL (Figure 1). The highest lovastatin yield occurred in the H20-tested strain. These results suggest that the lovastatin production capacity of wild Eurotium cristatum was in extreme variation. Among the six tested strains, the lovastatin yields of H11, H15, and H20 strains were twice than others. The H3 strain’s lovastatin yield was less than 2 μg/mL. All tested strains’ lovastatin yield was lower than Monascus and Aspergillus terreus (usually used for producing lovastatin).
At the same time, the content of Monascus and Aspergillus terreus was very low in Fuzhuan brick tea. Fuzhuan brick tea was considered a weight-reducing tea. And Eurotium cristatum is the dominant microorganism in Fuzhuan brick tea. Thus more lovastatin could enhance the lipid-lowering capacity of Fuzhuan brick tea. Hence, screening Eurotium cristatum with higher lovastatin yield was beneficial to develop new dark tea with excellent lipid-lowering capacity. The safety of a drink with lovastatin should be considered. Compared with wild Monascus (in the range of 10 μg/mL to 500 μg/mL in fermentation broth), the wild Eurotium cristatum strains produced lower lovastatin [6,8]. The red yeast rice fermented by Monascus has been made into many health foods, including red yeast rice powder, red yeast rice capsules, red yeast rice, and so on. And the lovastatin concentration in dark tea (always lower than 20 μg/g) was lower than in red yeast rice. Hence, dark tea is a safe drink for humans. A large number of studies showed that dark tea has a variety of health benefits [27].
Higher yields of statins were obtained with H11, H15, and H20 strains, whose lovastatin yields were more than 5 μg/mL in the fermentation broth. H15 and H20 strains were isolated from a Fuzhuan brick tea with well lipid-lowering capacity. The colony morphologies of strains producing lovastatin are exhibited in Figure 2. In contrast, the colony morphologies of the 6 tested strains were quite different, such as length of mycelium, smoothness, wrinkle, and so on. The H8, H11, H15, and H20 strains were smooth in the edge of the colony compared with H3 and H16 strains. The H3, H16, and H20 strains have wrinkles in the center of the colony. The H3 and H16 strain could produce a large brown pigment, and the H3 strain’s brown pigment was in the center of the colony. The mycelium length of the H11, H16, and H20 strains was longer than other strains. There was no one common characteristic in the colonial morphology of H11, H15, and H20 strains except brown pigment production. The pigment and lovastatin are polyketide metabolite. This suggested that the gene controlling colonial morphology did not correlate with gene-producing lovastatin. The length of colony diameter of strains producing lovastatin was larger than other strains which could not produce lovastatin. This indicated that these Eurotium cristatum strains could grow faster and accumulate more lovastatin.

3.2. Lovastatin Production in Mycelium, Fermentation Broth, and Propagule

After assessing the possibility of lovastatin production with strains, we focused on organ production by the H20 strain. As shown in Figure 3, the yields of lovastatin in mycelium, ascospore, and ascocarp were higher than in fermentation broth (more than twice). This means lovastatin was an intracellular product in Eurotium cristatum similar to Monascus and Aspergillus terreus [31,32,33]. Due to its lipid solubility and the small molecular weight of lovastatin, it will slowly release into the fermentation broth. The slow release rate resulted in lower lovastatin concentration in the fermentation broth. For the propagule of Eurotium cristatum, the 20 times higher yields of lovastatin in sexual propagule (ascospore and ascocarp) were observed compared with asexual propagule (conidium). These results suggested that lovastatin production was restrained in a hyperosmotic environment. Because conidium of Eurotium cristatum mainly grows under a hyperosmotic environment. At the same time, sexual propagule stored at Fuzhuan brick tea was beneficial to the accumulation of lovastatin. This was important for Fuzhuan brick tea to retain lipid-lowering metabolite due to the traditional Fuzhuan brick tea always contains a large yellow ascocarp. The more ascocarp in Fuzhuan brick induced higher lipid-lowering metabolite. Time was also beneficial to the accumulation of lovastatin. The lovastatin concentration in ascocarp separated from 6-year stored loose dark tea was higher (about 120%) than newly manufactured loose dark tea. These results indicated that the lipid-lowering functions of loose dark tea could enhance by extending storage time. The Fuzhuan brick tea should be stored in a ventilated and dry environment to inhibit the growth of other harmful microorganisms.

3.3. Influences of Fermentation Parameters on Lovastatin Production by H20 Strain

A detailed fermentation parameters study of lovastatin production by the H20 strain was carried out. The goal of this part was to obtain suitable conditions to produce more lovastatin by Eurotium cristatum strain. Initially, a time profiling of lovastatin production was studied using a PDA fluid medium. In the first few days, the concentration of lovastatin in the fermentation broth was linearly increased with increasing fermentation time (Figure 4). The yield of lovastatin passed through a maximum of 7 days of the experiments. The highest lovastatin yield was 7.24 μg/mL. After that time, the lovastatin concentration remained the same. These results indicated that lovastatin was no longer produced in the late fermentation process, maybe due to the consumption of nutrient substances in the fermentation broth, similar to Monascus producing lovastatin in the late fermentation process [34]. In the late fermentation process, they found that carbon and nitrogen source concentrations were reduced by more than 95%.
The production medium is PDA medium containing additional 4% glucose. To study the effect of different carbohydrate sources on lovastatin production, monosaccharide, disaccharide, and polysaccharide were used to replace glucose. As shown in Figure 5A, the carbohydrate source strongly affected lovastatin yields and was not dependent on the number of monosaccharides. For lovastatin production, glucose (monosaccharide) was better than the selected disaccharide. The type of disaccharide also affected lovastatin yields, and the lactose was unsuitable for producing more lovastatin. Those results suggested that Eurotium cristatum had a preference for carbohydrate sources to produce lovastatin.
In contrast to other carbohydrate sources, soluble starch was the best for lovastatin production. An approximately three-fold increase in lovastatin yield was observed compared to the experimental group, whose carbohydrate source was lactose. This experimental phenomenon may be because of its polysaccharide nature, which is difficult to absorb and utilize quickly. Hence, in a plateau of microbial growth, enough carbohydrate sources could be used to produce secondary metabolite [35].
The effect of the concentration of soluble starch on lovastatin production was also investigated. Under the lower concentration of soluble starch (less than 8%), the yield of lovastatin increased with an increase in its concentration. A total of 14.96 μg/mL lovastatin was produced by adding 8% soluble starch. This increase in yield was attributed to soluble starch addition, which helped to maintain the carbon-to-nitrogen (C:N) media ratio, and provided better conditions for lovastatin production. With further increasing soluble starch concentration, the yields of lovastatin will not increase. This change in lovastatin concentration may be attributed to the low solubility of soluble starch. For the concentration of soluble starch of more than 8%, the liquid fermentation medium was turbid.
Temperature affects enzymatic activity, then affects cell growth of microorganisms and accumulation of secondary metabolites. The optimum temperature for various secondary metabolite production and Eurotium cristatum growing could be different. Temperature is easy to control through a thermostat, so it is a useful fermentation factor to improve the yields of fermentation products. The highest lovastatin yield was achieved at 32 °C, and the lovastatin yield was 8.67 μg/mL. Further, an increase in temperature reduced lovastatin yield (Figure 6A). The enzyme’s activities that catalyze the synthesis of lovastatin depend on temperature. An increase in temperature may alter their spatial structure, which results in decreased lovastatin production. Eurotium cristatum is aerobe. Adequate oxygen is conducive to the growth of mycelium and the accumulation of metabolites. The dissolved oxygen in the fermentation broth can be controlled by adjusting the rotation speed. As shown in Figure 6B, a rotating culture flask produced the lovastatin, and the highest yield (10.27 μg/mL) was achieved at 120 r/min. Furthermore, an increase in rotation speed may destroy the mycelium because of the large shearing force or cause the formation of large mycelium pellets due to regular rotation. Those resulted in a decrease in lovastatin production.

4. Conclusions

In this study, several wild Eurotium cristatum strains in Anhua Fuzhuan brick tea were screened for lovastatin production. Among 20 wild-type strains, 6 Eurotium cristatum strains could form lovastatin. The highest lovastatin yield was observed in the H20 strain with a slow growth rate, no wrinkling, rough edges in the colony, and little brownness at the center. The colony morphologies type was not related to its lovastatin production capacity suggesting the gene-controlling colonial morphology did not correlate with gene-producing lovastatin. Among the organ of lovastatin production, the sexual propagule (ascospore and ascocarp) was the best. The asexual propagule (conidium) could produce little lovastatin. Traditional Fuzhuan brick tea and Jinhua Pu-erh tea always contain abundant ascocarp and little conidium, which is good for accumulating lipid-lowering substances in Fuzhuan brick tea. The sexual propagule stored in Fuzhuan brick tea was beneficial to the accumulation of lovastatin. The fermentation conditions to produce lovastatin by Eurotium cristatum H20 strain were also studied. The carbohydrate source strongly affects lovastatin yields, and soluble starch was the best. Compared with Monascus and Aspergillus terreus, a little lovastatin was detected in wild Eurotium cristatum strains. This will limit the accumulation of lovastatin in Fuzhuan brick tea. To increase lipid-lowering substance in Fuzhuan brick tea, the breeding of Eurotium cristatum with higher lovastatin yields should be carried out in the future.

Author Contributions

Conceptualization, T.L. and Z.L. (Zhonghua Liu); methodology, Z.L. (Zhanjun Liu) and P.L.; validation, J.L. and Y.Z.; formal analysis, T.L. and Z.L. (Zhanjun Liu); investigation, J.L. and T.L.; resources, P.L., Y.Z. and J.L.; writing—original draft preparation, T.L. and Z.L. (Zhanjun Liu); writing—review and editing, T.L. and Z.L. (Zhonghua Liu); visualization, J.L. and T.L.; project administration, Z.L. (Zhonghua Liu) and P.L.; project administration, P.L. All authors have read and agreed to the published version of the manuscript.

Funding

This work was supported by the Hunan Provincial Key Lab of Dark Tea and Jin-hua (2016TP1022), the Research Foundation of Education Bureau of Hunan Province (22B0785), the Natural Science Foundation of regional joint fund project of Hunan Province (2023JJ50338).

Data Availability Statement

The data presented in this study are available on request from the corresponding author.

Conflicts of Interest

The authors declare no conflict of interest.

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Figure 1. Lovastatin yields in different screened Eurotium cristatum strains. The lovastatin concentration was detected in the fermentation broth. Strains were fermented on a PDA fluid medium at 28 °C for 8 days. The error bars are standard deviations.
Figure 1. Lovastatin yields in different screened Eurotium cristatum strains. The lovastatin concentration was detected in the fermentation broth. Strains were fermented on a PDA fluid medium at 28 °C for 8 days. The error bars are standard deviations.
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Figure 2. Colony morphologies of various Eurotium cristatum strains producing lovastatin. Strains were grown for 9 days on PDA medium at 28 °C.
Figure 2. Colony morphologies of various Eurotium cristatum strains producing lovastatin. Strains were grown for 9 days on PDA medium at 28 °C.
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Figure 3. Lovastatin production in mycelium, fermentation broth, and propagule of the Eurotium cristatum H20 strain. Strains were fermented at 28 °C for 8 days. The propagule includes sexual propagule (ascospore and ascocarp) and asexual propagule (conidium). Six-year ascopore and ascocarp were separated from loose dark tea stored in a ventilated and dry environment for 6 years. The error bars are standard deviations.
Figure 3. Lovastatin production in mycelium, fermentation broth, and propagule of the Eurotium cristatum H20 strain. Strains were fermented at 28 °C for 8 days. The propagule includes sexual propagule (ascospore and ascocarp) and asexual propagule (conidium). Six-year ascopore and ascocarp were separated from loose dark tea stored in a ventilated and dry environment for 6 years. The error bars are standard deviations.
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Figure 4. Time profile of lovastatin production by Eurotium cristatum H20 strain on PDA fluid medium at 28 °C. The lovastatin concentration was detected in the fermentation broth. The error bars are standard deviations.
Figure 4. Time profile of lovastatin production by Eurotium cristatum H20 strain on PDA fluid medium at 28 °C. The lovastatin concentration was detected in the fermentation broth. The error bars are standard deviations.
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Figure 5. The influences of additional carbohydrate sources (A) and concentration of soluble starch (B) on lovastatin production. Strains were fermented at 28 °C for 8 days. The lovastatin concentration was detected in Eurotium cristatum H20 strain fermentation broth. The error bars are standard deviations.
Figure 5. The influences of additional carbohydrate sources (A) and concentration of soluble starch (B) on lovastatin production. Strains were fermented at 28 °C for 8 days. The lovastatin concentration was detected in Eurotium cristatum H20 strain fermentation broth. The error bars are standard deviations.
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Figure 6. The influences of temperature (A) and rotation speed (B) on lovastatin production. Strains were fermented for 8 days. The lovastatin concentration was detected in Eurotium cristatum H20 strain fermentation broth. The error bars are standard deviations.
Figure 6. The influences of temperature (A) and rotation speed (B) on lovastatin production. Strains were fermented for 8 days. The lovastatin concentration was detected in Eurotium cristatum H20 strain fermentation broth. The error bars are standard deviations.
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Li, T.; Liu, Z.; Li, J.; Zheng, Y.; Liu, Z.; Ling, P. Lovastatin Production by Wild Eurotium cristatum Isolated from Fuzhuan Brick Tea Produced Using Forest Resources in Auhua. Forests 2023, 14, 1409. https://doi.org/10.3390/f14071409

AMA Style

Li T, Liu Z, Li J, Zheng Y, Liu Z, Ling P. Lovastatin Production by Wild Eurotium cristatum Isolated from Fuzhuan Brick Tea Produced Using Forest Resources in Auhua. Forests. 2023; 14(7):1409. https://doi.org/10.3390/f14071409

Chicago/Turabian Style

Li, Taotao, Zhanjun Liu, Jun Li, Yajun Zheng, Zhonghua Liu, and Peixue Ling. 2023. "Lovastatin Production by Wild Eurotium cristatum Isolated from Fuzhuan Brick Tea Produced Using Forest Resources in Auhua" Forests 14, no. 7: 1409. https://doi.org/10.3390/f14071409

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