Lactobacillus mucilaginosus FW03 and application thereof
By fermenting noni juice with Lactobacillus mucilaginosus FW03, the rancidity problem of noni juice was solved, the total phenol and total flavonoid contents were increased, the antioxidant capacity was enhanced, the flavor was improved, and the market application prospects were expanded.
Patent Information
- Application Number
- CN202510852548.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-24
- Publication Date
- 2025-09-23
AI Technical Summary
After ripening, noni juice becomes soft and juicy with a strong rancid smell, which makes it difficult for consumers to accept it. Existing improvement methods have failed to effectively increase its flavor and active ingredient content.
Noni juice was fermented with Lactobacillus mucilaginosus FW03 at an inoculation rate of 10% and fermented at 37°C for 7 days to improve the flavor of the juice and increase the content of active ingredients.
It significantly increased the total phenol and total flavonoid content of noni juice, enhanced the antioxidant capacity, improved the flavor of the juice, made it softer, and increased consumer acceptance.
Smart Images
Figure CN120682995A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of fermentation engineering and relates to fermented Lactobacillus mucilaginosus FW03 and application thereof. Background Art
[0002] Noni, also known as Morinda officinalis, belongs to the genus Morinda in the Rubiaceae family. It is a perennial evergreen plant that grows in tropical and subtropical regions, mostly as a small tree or shrub. Noni has broad, oval leaves and grenade-shaped fruit, which range in color from green to yellow to nearly white at harvest. A strong, rancid odor characterizes the ripe fruit, which is a light, dark yellow or white color. The flesh is juicy and has a bitter or astringent taste. Noni fruit (Morinda citrifolia) is rich in nutritional value, containing over 200 active substances such as coumarins, flavonoids, and vitamins. Its unique health benefits have made it a hot topic for research and development. However, upon ripening, the fruit becomes soft and juicy, inevitably exhibiting a strong rancid odor, making it unacceptable to many consumers.
[0003] To improve the flavor of pure noni juice, many companies are adding popular fruits like blueberries, apples, and grapes to the original noni juice, improving the flavor and expanding the market for noni products. To improve the flavor and quality of noni products, noni juice can be processed through microbial inoculation and fermentation. This fermentation process has significantly improved the quality and flavor of noni juice. Summary of the Invention
[0004] Based on the above, the present invention aims to provide fermentative Lactobacillus mucilaginosus FW03 and its application. By screening out a strain of fermentative Lactobacillus mucilaginosus FW03 with good growth performance, it is used for noni fermentation to increase the active ingredient content of fermented noni juice, improve the unpleasant flavor of fermented noni juice, increase the floral ester compounds in fermented noni juice, make the fermented noni juice flavor fresh and soft, improve consumer acceptability, and expand market application prospects.
[0005] The technical solution adopted by the present invention to achieve the technical purpose is:
[0006] The present invention provides a strain of fermented mucus lactobacillus (Limosilactobacillus fermentum) FW03, characterized in that it has been deposited in the Guangdong Provincial Microbiological Collection Center (GDMCC) on May 30, 2025, with a preservation address of 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, with a preservation number of GDMCC No: 66444 and a classification name of Limosilactobacillus fermentum.
[0007] The present invention also provides a noni fruit fermentation agent containing the fermented mucilaginous Lactobacillus FW03.
[0008] The present invention further provides a method for fermenting noni fruit, comprising: inoculating the fermented Lactobacillus mucilaginosus FW03 into pasteurized noni fruit pulp at a rate of 10%, and fermenting at 37° C. for 7 days.
[0009] The present invention further provides the use of the fermented Lactobacillus mucilaginosus FW03 in the fermentation of noni fruit.
[0010] Furthermore, Lactobacillus mucilaginosus FW03 was inoculated into pasteurized noni pulp at an inoculum amount of 10%, and fermented at 37° C. for 7 days.
[0011] Furthermore, fermenting noni pulp with Lactobacillus mucilaginosus FW03 can improve the flavor of noni juice.
[0012] Furthermore, fermenting noni pulp with Lactobacillus mucilaginosus FW03 can enhance the DPPH free radical scavenging ability of noni fermented juice.
[0013] Furthermore, fermenting noni pulp with Lactobacillus mucilaginosus FW03 can enhance the ABTS free radical scavenging ability of noni fermented juice.
[0014] Furthermore, fermenting noni pulp with Lactobacillus mucilaginosus FW03 can significantly increase the content of total phenols in the fermented noni juice.
[0015] Furthermore, fermenting noni pulp with Lactobacillus mucilaginosus FW03 can significantly increase the content of total flavonoids in the fermented noni juice.
[0016] The beneficial effects of the present invention are:
[0017] The fast-growing Lactobacillus fermentum FW03 strain screened by the present invention can improve the taste of fermented Noni juice, increase the flavor compounds in the juice, impart fragrance to the juice, and soften the flavor. Furthermore, fermenting Noni juice with the Lactobacillus fermentum FW03 significantly increases the total phenolic and flavonoid content of the juice and enhances the antioxidant activity of the juice. BRIEF DESCRIPTION OF THE DRAWINGS
[0018] Figure 1 This is a growth curve of the fermentation Lactobacillus mucilaginosus FW03 of the present invention;
[0019] Figure 2This is a phylogenetic tree diagram of the fermentation Lactobacillus mucilaginosus FW03 of the present invention;
[0020] Figure 3 This is a radar chart for the sensory evaluation of the fermented Lactobacillus mucilaginosus FW03 of the present invention, the commercially fermented Lactobacillus mucilaginosus HH-LF39 and the naturally fermented Noni juice. DETAILED DESCRIPTION
[0021] The present invention will be described in detail below with reference to specific embodiments. The following specific embodiments will help those skilled in the art to further understand the present invention, but will not limit the present invention in any form.
[0022] Example 1 Isolation and identification of fermentative Lactobacillus mucilaginosus FW03
[0023] Fermented mucus Lactobacillus FW03 is a lactic acid bacterium isolated and purified from Hainan's specialty fermented sour melon. The strain growth curve of the screened fermented mucus Lactobacillus FW03 was determined, and the 16SrRNA gene of the strain was sequenced and submitted to NBCI for homologous comparative identification.
[0024] like Figure 1 The 24h growth curve of Lactobacillus fermentatus FW03 is shown. It enters the logarithmic growth phase at 2h, reaches the end of the logarithmic growth phase at 10h, and then enters the stable phase. The bacterial density reaches the maximum value at the end of the logarithmic phase and enters the growth decline phase at 20h. The bacterial density is relatively stable, indicating good fermentation stability. The isolated and screened strain FW03 was identified, and its nucleotide sequence is shown in SEQ ID NO.1. Further comparison and analysis with NCBI were performed to screen strains with high consistency with the 16SrRNA sequence of the FW03 strain, and a phylogenetic evolutionary tree was made, as shown in the following figure: Figure 2 The similarity between strain FW03 and Lactobacillus fermentans was 100%, and it was speculated that the strain was Lactobacillus fermentans.
[0025] The isolated and purified fermented mucus Lactobacillus FW03 was deposited in the Guangdong Provincial Microbiological Collection Center (GDMCC) on May 30, 2025. The deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. The deposit number is GDMCC No: 66444, and the classification name is: Limosilactobacillus fermentum.
[0026] Example 2 Fermentation of Noni Juice by Lactobacillus mucilaginosus FW03
[0027] The specific experimental groups of the present invention are set as follows: FW03 fermentation group, HH-LF39 fermentation group, and natural fermentation group.
[0028] Raw Materials and Fruit Puree Preparation: Fresh noni fruit is harvested from a noni fruit farm in Wanning City, Hainan Province. Noni plants with identical bases are selected, and fresh fruit is harvested at 8-9 maturity. Fruit that is free of mechanical damage and rot is selected, washed thoroughly with running water, and air-dried. The fruit is then cut into small pieces, crushed and pressed into a pulp using a pulper. The pulp is then packaged in sterilized fermentation tanks, pasteurized, and cooled for later use.
[0029] Activation: Inoculate a 2% inoculum of Lactobacillus mucilaginosus FW03 into MRS liquid medium and incubate at 37°C for two generations until the logarithmic growth phase. The activated culture was centrifuged at 8000 rpm for 10 minutes, the supernatant discarded, and the cells washed three times with sterile saline. The cells were harvested and resuspended in the same volume of 0.9% sterile saline. Commercial Lactobacillus mucilaginosus HH-LF39 (purchased from Shaanxi Ruimao Biotechnology Co., Ltd.) was prepared as described above.
[0030] Inoculation and fermentation: The activated fermented Lactobacillus mucilaginosus FW03 and commercial fermented Lactobacillus mucilaginosus HH-LF39 were inoculated at a rate of 10% (OD 600nm =1.2) were inoculated into pasteurized Noni pulp and fermented at 37°C for 7 days. Naturally fermented Noni juice was used as a control.
[0031] 1.1 Determination of total phenols and flavonoids in fermented noni juice
[0032] The total phenol content in fermented noni juice was determined using the Folin-phenol method. A stock solution of gallic acid was prepared at a concentration of 200 μg / mL. This solution was diluted to produce a series of standard solutions with concentrations ranging from 0 to 200 μg / mL. 50 μL of the diluted fermented juice sample or standard solution was placed in an EP tube. 50 μL of the Folin-phenol colorimetric reagent was added and reacted for 10 minutes. 200 μL of a 20% Na₂CO₃ solution was then added and reacted for 20 minutes. The absorbance was then measured at 765 nm. The total flavonoid content in fermented noni juice was determined using the sodium nitrite-aluminum chloride method. A stock solution of rutin was prepared at a concentration of 1 mg / mL. This solution was diluted to produce a series of standard solutions with concentrations ranging from 150 to 900 μg / mL. Take 25 μL of sample or standard solution, add 110 μL of 0.066 mol / L sodium nitrite solution, let it stand for 5 minutes, then add 15 μL of 0.5 mol / L aluminum chloride solution, let it stand for 6 minutes, then add 100 μL of 0.5 mol / L sodium hydroxide solution, and react for 10 minutes. Measure the absorbance at 510 nm.
[0033] 1.2 DPPH free radical scavenging activity and ABTS of fermented noni juice + 1.2.1 Determination of free radical scavenging ability DPPH free radical scavenging ability
[0034] Prepare a 0.1mmol / L DPPH solution and dilute to volume with methanol. Take 50μL of sample solution, add 400μL of DPPH working solution, mix thoroughly, and react at room temperature in the dark for 30 minutes. Measure the absorbance at a wavelength of 517nm. Use trolox as a positive control and draw a standard curve. The formula for calculating DPPH free radical scavenging rate is:
[0035]
[0036] Where: A i : Sample absorbance value; A j : absorbance value when methanol replaces DPPH; A c : Absorbance value when methanol is used instead of sample.
[0037] 1.2.2ABTS + Determination of free radical scavenging ability
[0038] Mix equal volumes of 7mmol / L ABTS solution and 2.45mmol / L potassium persulfate solution. Incubate in the dark at room temperature for 16 hours. Dilute the newly prepared ABTS solution with distilled water to an absorbance of 0.70±0.02 at 734nm. Add 50μL of sample solution to 400μL ABTS working solution, mix thoroughly, and incubate in the dark at room temperature for 30 minutes. Measure the absorbance at 734nm. Use trolox as a positive control and plot a standard curve. The formula for calculating the free radical scavenging ability of ABTS is as follows:
[0039]
[0040] Where: A i : Sample absorbance value; A j : absorbance value when methanol replaces ABTS; A c : Absorbance value when methanol is used instead of sample.
[0041] Table 1 Total phenols, total flavonoids and antioxidant activity of noni juice fermented by different strains
[0042]
[0043] The results are shown in Table 1. The total phenol content of noni juice fermented with Lactobacillus mucilaginosus FW03 was 0.74 mg / mL, significantly higher than that of naturally fermented juice and commercially fermented juice with Lactobacillus mucilaginosus HH-LF39. Therefore, fermentation with Lactobacillus mucilaginosus FW03 significantly increased the total phenol content of the fermented juice, and the fermentation efficiency was superior to that of commercially fermented juice with Lactobacillus mucilaginosus HH-LF39. The total flavonoid content of noni juice fermented with Lactobacillus mucilaginosus FW03 was 0.49 mg / mL, higher than that of the naturally fermented juice. However, the total flavonoid content of noni juice fermented with commercially fermented Lactobacillus mucilaginosus HH-LF39 was 0.28 mg / mL, indicating a decrease in total phenolic content. This indicates that the total flavonoid content of noni juice fermented with FW03 was significantly increased. The antioxidant capacity of the two strains in noni juice fermentation was significantly different. The DPPH radical scavenging capacity of the juice fermented with FW03 was 89.65%, while that of the juice fermented with HH-LF39 was 86.88%, both exceeding that of the naturally fermented juice. Overall, fermentation with Lactobacillus mucilaginosus FW03 improved the DPPH radical scavenging capacity of juice, and the fermentation effect was superior to that of the commercial Lactobacillus mucilaginosus HH-LF39. The ABTS+ radical scavenging capacity of juice fermented with FW03 was 90.94%, while that of juice fermented with the commercial Lactobacillus mucilaginosus was 86.22%. FW03 fermentation in noni fruit pulp was superior to that of the commercial Lactobacillus mucilaginosus HH-LF39, effectively improving the antioxidant capacity of the juice.
[0044] Example 3 Sensory Evaluation of Fermented Noni Juice
[0045] A sensory panel of 10 trained sensory assessors (aged 20-35) evaluated the fermented noni juices. The panelists objectively assessed the texture, color, and flavor of the samples based on pre-defined criteria. The scores were divided into nine levels, ranging from 0 (very weak) to 9 (very strong), based on the strength of the characteristic. The scoring criteria are shown in Table 2.
[0046] Table 2 Sensation scoring criteria
[0047]
[0048]
[0049] Depend on Figure 3It can be seen that the tissue states of the three fermented juices are not much different, and the color of the juice fermented by Lactobacillus mucilaginosus FW03 is not much different from that of the naturally fermented juice, indicating that fermentation by Lactobacillus mucilaginosus FW03 can maintain the color of the juice. Among the sensory evaluation indicators, sourness reflects the taste reaction related to citric acid in the juice. Among the three fermented juices, the sourness of the naturally fermented group was stimulating, while the sourness of the noni juice fermented by Lactobacillus mucilaginosus FW03 was relatively mild, indicating that FW03 is more in line with the public's preference for sour-sweet balance in acidity regulation; sweetness reflects the taste reaction related to fructose, glucose and sucrose in the juice. Among the three groups of fermented juices, the juice fermented by Lactobacillus mucilaginosus FW03 had the strongest sweetness, while the juice of the HH-LF39 fermentation group and the naturally fermented group had weaker sweetness; the cheese flavor (sour and smelly smell) of the juice decreased after inoculation and fermentation, and the cheese flavor of the FW03 fermentation group decreased significantly, indicating that inoculation and fermentation can reduce the unpleasant odor of the fermented juice; the fermentation flavor reflects the flavor related to acids and alcohols in the fermented fruit brought to the juice by the fermentation of the bacteria, reflecting the strength of the fermentation degree. The FW03 fermentation group scored high, while the HH-LF39 and natural fermentation groups scored lower. This reflects the FW03 fermentation group's richer lactic acid and other unique flavors, which intensify the fermentation flavor. Aroma indicators are characterized by a refreshing fruity aroma. The FW03 fermentation group scored the highest, indicating that the FW03 fermentation group improved the aroma of the fermented juice. The FW03 fermentation group scored the highest, thanks to the synergistic advantages of texture, fermentation flavor, and sweetness, resulting in a rich and stable flavor profile.
[0050] HS-SPME-GC-MS analysis of flavor compounds in noni juice fermented with Lactobacillus mucilaginosus FW03 revealed a decrease in caproic acid and octanoic acid content in the juice after FW03 fermentation. FW03 fermentation also softened the cheesy and pungent sourness of the juice, softening the acidity and reducing the heavy, fermented flavor for a lighter flavor. Esters are the primary source of fruity and floral aromas. Compared to natural fermentation, FW03 fermentation significantly increased the levels of six volatile aroma components. The significantly higher isoamyl phenylacetate content in the juice fermented with FW03 was 1120.63 μg / kg, while the naturally fermented juice contained 567.46 μg / kg of isoamyl phenylacetate. This significantly increased content imparts a richer floral aroma to the juice. The linalool content in FW03-fermented juice was 51.63 μg / kg, significantly higher than that in naturally fermented juice. Its lemon and rose aromas significantly enhanced the floral aroma of the fermented juice, becoming the primary aroma-active substances. The newly generated α-terpineol and n-octanol in FW03-fermented noni juice enriched the aroma of the fermented juice to varying degrees. α-terpineol, with its minty and aniseed notes, added a refreshing touch. FW03-fermented juice also produced two aldehydes, benzaldehyde and nonanal, which imparted almond and caramel flavors. The floral, grassy, and lemony notes of nonanal further enhanced the refreshing effect.
[0051] Table 3 Contents of flavor substances and aroma activity values of different noni fermented juices
[0052]
[0053] Note: “-” means not detected
[0054] Obviously, the above embodiments of the present invention are merely examples to more clearly illustrate the present invention, and are not limitations on the implementation methods of the present invention. For ordinary technicians in the relevant field, other different forms of changes or modifications can be made based on the above description. It is impossible to enumerate all implementation methods here. All obvious changes or modifications derived from the technical solution of the present invention are still within the scope of protection of the present invention.
Claims
1. A strain of Lactobacillus fermentum FW03, characterized in that: It was deposited in Guangdong Provincial Microbiological Collection Center on May 30, 2025. The deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City. The deposit number is GDMCC No: 66444. The classification name is: Limosilactobacillus fermentum.
2. A Noni fruit fermentation agent, characterized in that: Contains the fermented Lactobacillus mucilaginosus FW03 according to claim 1.
3. A method for fermenting Noni fruit, characterized in that: include: The fermented Lactobacillus mucilaginosus FW03 according to claim 1 was inoculated into pasteurized Noni pulp at an inoculum rate of 10%, and fermented at 37° C. for 7 days.
4. Application of the fermented Lactobacillus mucilaginosus FW03 according to claim 1 in Noni fruit fermentation.
5. The use according to claim 4, characterized in that Lactobacillus mucilaginosus FW03 was inoculated into pasteurized noni pulp at a 10% inoculum rate and fermented at 37°C for 7 days.
6. The use according to claim 5, characterized in that Fermenting noni pulp with Lactobacillus mucilaginosus FW03 can improve the flavor of noni juice.
7. The use according to claim 4, characterized in that Fermenting noni pulp with Lactobacillus mucilaginosus FW03 can enhance the DPPH free radical scavenging ability of noni fermented juice.
8. The use according to claim 4, characterized in that Fermenting noni pulp with Lactobacillus mucilaginosus FW03 can enhance the ABTS free radical scavenging ability of noni fermented juice.
9. The use according to claim 4, characterized in that Fermenting noni pulp with Lactobacillus mucilaginosus FW03 can significantly increase the content of total phenols in the fermented noni juice.
10. The use according to claim 4, characterized in that Fermenting noni pulp with Lactobacillus mucilaginosus FW03 can significantly increase the content of total flavonoids in the fermented noni juice.