CN114369558B - Serratia marcescens and application thereof in naringinase production - Google Patents
Serratia marcescens and application thereof in naringinase production Download PDFInfo
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Abstract
The invention discloses serratia marcescens and application thereof in naringinase production, and belongs to the technical field of microorganisms. The strain is named as serratia marcescens (A)Serratia marcescens) C10, deposited at No. 59 of the great college, Jie Lianlu 100, Calif., Guangdong province, in Guangzhou, Youzhou, 1/19, 2022, with the deposit number GDMCC No. 62223. The strain is a novel strain capable of producing naringinase, can efficiently catalyze and hydrolyze naringin, has naringin hydrolysis efficiency of about 90 percent, and has good activity and stability. Therefore, the serratia marcescens C10 has important application potential in debittering processing and flavor improvement of different citrus juices.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to serratia marcescens and application thereof in naringinase production.
Background
With the rapid development of food and beverage processing industry in China and the continuous improvement of requirements of people on healthy diet quality, drinks rich in flavonoid compounds such as orange juice are widely favored by consumers, however, the orange juice is rich in a large amount of flavonoid compounds such as hesperidin and naringin, so that the bitter taste is strong, and the taste and the sales are seriously influenced. On the one hand, naringinase mainly contains two enzymes, one is alpha-L-rhamnosidase which can hydrolyze bitter naringin into rhamnose and naringenin-7-O glucoside, and the second is beta-D-glucosidase which can hydrolyze naringenin-7-O glucoside into tasteless naringenin and glucose. Eliminate bitter taste of fruit juice and improve its nutritional components. The products of hesperetin or naringenin and the like after the action of the enzyme are key precursors for producing the high-value sweetener neohesperidin dihydrochalcone; on the other hand, the enzyme can be used for debitterizing orange juice, can promote the fragrance of wine, eliminate hesperidin, naringin and other crystals in the orange juice and improve the antioxidant activity of the citrus juice. However, the stability of the natural free naringinase is not high at present, and the industrial production and processing conditions cannot be met.
The glycosidic bond in flavonoid compounds such as hydrolyzed hesperidin and naringin mainly has two chemical and biological methods, wherein the microbial enzyme technology overcomes the defects of serious pollution and the like of a chemical acid hydrolysis process, and the enzyme method debitterizing has the advantages of strong specificity, good debitterizing effect, simple operation, mild debitterizing condition, good debitterizing effect, no damage to other nutrient components and flavor in the fruit juice, safety and reliability of the enzyme method and the like. Therefore, the microbial enzyme method becomes the most widely applied fruit juice debittering technology at home and abroad at present.
It is reported in literature that naringinase is mainly derived from natural fungi such as Aspergillus niger, Penicillium and Aspergillus oryzae, and a small amount of naringinase is derived from bacteria and yeasts such as Staphylococcus xylosus and Cryptococcus laurentii. Wherein, the enzyme properties and the application range of the enzyme of the microorganism producing naringinase from different sources are different. Therefore, the screening of the bacterial strain capable of producing the naringinase with high activity and stability has important practical significance.
Disclosure of Invention
In order to solve the related problems, the invention mainly aims to provide a serratia marcescens strain.
Another object of the present invention is to provide the use of the above-mentioned Serratia marcescens for the production of naringinase.
It is still another object of the present invention to provide a biological agent having naringinase activity.
In order to achieve the purpose, the invention adopts the following technical scheme:
a strain of serratia marcescens, named serratia marcescens ()Serratia marcescens) C10, deposited at 19.1.2022 at the Guangdong province microbial culture Collection (GDMCC) of building 59 of Zhou Lu 100, Pieli, Calif., Guangzhou, Guangdong province, with the deposit number GDMCC No. 62223.
The serratia marcescens C10 is applied to the production of naringinase.
Further, the naringinase consists of alpha-L-rhamnosidase and beta-D-glucosidase.
A method for producing naringinase by using the serratia marcescens C10 is realized by separating and purifying cell disruption liquid of serratia marcescens C10; the method specifically comprises the following steps:
(1) inoculating serratia marcescens C10 to a solid culture medium for culture, and then inoculating to a liquid culture medium for culture to obtain a fermentation liquid;
(2) centrifugally separating the fermentation liquor, discarding the supernatant, resuspending the precipitate, and performing ultrasonic disruption to obtain a cell disruption solution;
(3) separating and purifying the cell disruption solution to obtain naringinase.
Further, the inoculation in the solid culture medium in the step (1) is carried out for 1-3 days at the temperature of 28-32 ℃ and at the speed of 150-200 r/min; preferably, the culture is carried out at 30 ℃ and 180r/min for 3 days.
Further, the solid medium described in step (1) contains per 100 mL: 0.15-0.25 g of naringin, 0.05-0.15 g of ammonium sulfate, 0.01-0.1 g of magnesium sulfate, 0.05-0.15 g of dipotassium hydrogen phosphate, 0.01-0.03 g of calcium chloride, 0.02-0.04 g of yeast, 0.02-0.04 g of beef extract, 0.5-1.5 g of agar powder and the balance of water; preferably contains: naringin 0.2 g, ammonium sulfate 0.1 g, magnesium sulfate 0.05 g, dipotassium hydrogen phosphate 0.1 g, calcium chloride 0.02 g, yeast 0.03 g, beef extract 0.03 g, agar powder 1g and water in balance.
Further, the conditions of inoculating the strain in the liquid culture medium and culturing in the step (1) are that the strain is cultured for 1-3 days at the temperature of 28-32 ℃ and at the speed of 150-200 r/min; preferably, the culture is carried out at 30 ℃ and 180r/min for 1 day.
Further, the liquid medium described in step (1) contains per 100 mL: 0.15-0.25 g of naringin, 0.05-0.15 g of ammonium sulfate, 0.01-0.1 g of magnesium sulfate, 0.05-0.15 g of dipotassium hydrogen phosphate, 0.01-0.03 g of calcium chloride, 0.02-0.04 g of yeast, 0.02-0.04 g of beef extract and the balance of water; preferably contains: naringin 0.2 g, ammonium sulfate 0.1 g, magnesium sulfate 0.05 g, dipotassium hydrogen phosphate 0.1 g, calcium chloride 0.02 g, yeast 0.03 g, beef extract 0.03 g, and water in balance.
Further, the inoculation amount of the inoculation in the liquid culture medium in the step (1) is 1-2%.
Further, the centrifugation condition in the step (2) is 8000-12000 r/min, and the centrifugation is carried out for 8-12 min at the temperature of 3-5 ℃; preferably 10000 r/min, 4 ℃ centrifugation for 10 min.
Further, the reagent used in the resuspension in step (2) is PBS buffer with pH8.0 ± 0.5, and the amount is 1g: 10mL, preferably, in terms of the precipitation reagent =1g: 5-15 mL.
Further, the ultrasonic condition in the step (2) is 150-250W, crushing is carried out for 2 s, stopping for 2 s, and the time is 10 min; preferably 200W, for 2 s of crushing and 2 s of stopping, for 10 min.
Naringinase is prepared by the above method.
A biological agent with naringinase activity comprises thallus or cell disruption solution of Serratia marcescens C10.
Further, the cell disruption solution is prepared by the steps (1) and (2) of the method.
The serratia marcescens C10, the naringinase or the biological preparation with the naringinase activity are applied to the processing treatment of citrus juice and/or the biotransformation of flavonoids compounds.
Further, the flavonoid compounds comprise hesperidin, naringin and dihydrochalcone derivatives thereof.
Further, the application comprises the following steps: mixing a flavonoid compound, a cell disruption solution of serratia marcescens C10 and a citric acid buffer solution, reacting for 25-35 min at 45-55 ℃, and inactivating for 8-12 min at 95-100 ℃; preferably, after 30min of reaction at 50 ℃, inactivation is carried out for 10min at 100 ℃.
Further, the citric acid buffer solution is 45-55 mmol/L of citric acid buffer solution with the pH value of 6 +/-0.2; preferably 50 mmol/L citrate buffer pH 6.
Further, the proportion of the flavonoid compound, the cell disruption solution of the serratia marcescens C10 and the citric acid buffer solution is 4-6 mg, 1mL and 1 mL; preferably 5 mg: 1 mL: 1 mL.
Compared with the prior art, the invention has the following advantages and effects:
the strain serratia marcescens C10 can quickly and efficiently produce naringinase, the efficiency of naringinase catalyzing and hydrolyzing naringin and derivatives thereof such as dihydrochalcone reaches about 90%, and the activity and the stability of the produced naringinase are good. Thus, Serratia marcescensSerratia marcescensThe naringinase produced by C10 has good fruit juice debittering effect, can be applied to processing debittering and flavor improvement of various citrus fruit juices, and has good application prospect.
Drawings
FIG. 1 is a colony morphology of Serratia marcescens C10.
FIG. 2 is a cell morphology of Serratia marcescens C10 under microscope.
FIG. 3 is a graph showing a standard ρ NP method.
FIG. 4 is a graph showing the results of crude enzyme activity measurement.
FIG. 5 is a graph showing the effect of Serratia marcescens C10 in hydrolyzing Naringin Dihydrochalcone (NDC).
Detailed Description
The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
Example 1: serratia marcescensSerratia marcescensIsolation and characterization of C10
Samples were taken from soil at the three-red-grass generation plantation site (N25 DEG 41 '29'; E110 DEG 35 '44') of Guilin, Guangxi. 10 g of mildewed soil and shaddock peel are taken to be put into 50 mL of sterile water and shaken for half an hour at 30 ℃ and 180 r/min. Then 500. mu.L of the supernatant was taken in 100mL of enrichment medium. Shaking at 30 ℃ for 3 days at 180 r/min. Carrying out gradient dilution to 10-5Spreading 10 μ L of the suspension on a plate, culturing at 30 deg.C for 3 days, spraying diethylene glycol alkali solution on the surface of the plate, and streaking for 3 days when a transparent ring appears on the colony. And finally, fermenting and culturing the single colony in a liquid culture medium for 3 days at 30 ℃ for 180r/min, and performing subsequent glycerin tube storage to obtain a new strain. The culture medium comprises liquid culture medium (g/100 mL) containing naringin 0.2, ammonium sulfate 0.1, magnesium sulfate 0.05, dipotassium hydrogen phosphate 0.1, calcium chloride 0.02, yeast 0.03, and beef extract 0.03; solid medium (g/100 mL): naringin 0.2, ammonium sulfate 0.1, magnesium sulfate 0.05, dipotassium hydrogen phosphate 0.1, calcium chloride 0.02, yeast 0.03, beef extract 0.03, and agar powder 1; sterilizing at 121 deg.C under high temperature and high pressure for 15 min.
Morphological feature analysis: as shown in figure 1 and figure 2, the bacterial strain is cultured on a solid medium at 30 ℃ for 3 days, and the colony morphology is characterized in that: the bacterial colony is beige, irregular edge, smooth surface, luster, flat and opaque center, and the diameter of the bacterial colony is 1-2.5 mm. The cells are in a shape of a nearly spherical short rod; the cell morphology characteristics of the compound are as follows: the size of the flagellum is 0.5-1.0 μm, and the flagellum is generated.
Physiological and biochemical characteristics analysis, as shown in table 1:
16S rDNA sequence analysis: extracting genome DNA of the strain by using a bacterial genome extraction kit, amplifying by using bacterial 16S rRNA gene universal primers 27F/1492R (27F: 5'-AGAGTTTGATCATGGCTCAG-3' (SEQ ID NO. 2) and 1492R: 5'-TACGGTTACCTTGTTACGACTT-3' (SEQ ID NO. 3)) to obtain a PCR product and sequencing, wherein the sequence is shown as SEQ ID NO.1, the sequencing result is analyzed by BLAST comparison, and in the comparison result of the bacterial strain 16S rDNA region and NCBI database, the sequencing result is compared with NCBI databaseSerratia marcescens The similarity is highest, and the matching degree reaches 99.86%.
Based on the analysis results, the strain is preliminarily judged to be serratia marcescensSerratia marcescensThis strain was named Serratia marcescens (A), (B), (C)Serratia marcescens) C10, and deposited at No. 59 building of the Zhou Lu Miao No. 100, Rehdea, Calif., Guangdong province at 19.1.1.2022, with the deposit number of GDMCC No. 62223.
Example 2: serratia marcescensSerratia marcescens Confirmation of C10 naringinase production and enzyme activity determination
1. Preparation of crude enzyme solution
Inoculating the preserved glycerol strain on a solid culture medium, culturing at 30 deg.C for 3 days, inoculating in a liquid culture medium, and culturing at 30 deg.C for 3 days at 180 r/min. Centrifuging the fermentation liquid at 4 ℃ for 10min at 10000 r/min for 10min, discarding the supernatant, adding 10mL of 50 mM PBS buffer solution with pH8.0, centrifuging at 4 ℃ for 5min at 10000 r/min, repeating the operation twice, discarding the supernatant, and finally adding PBS buffer solution according to a ratio of 1:10 (g: mL) for carrying out ultrasonic disruption. Ultrasonic crushing conditions: 200W, crushing for 2 s, stopping for 2 s, and taking 10 min.
Solid medium (g/100 mL): naringin 0.2, ammonium sulfate 0.1, magnesium sulfate 0.05, dipotassium hydrogen phosphate 0.1, calcium chloride 0.02, yeast 0.03, beef extract 0.03, and agar powder 1;
liquid medium (g/100 mL): naringin 0.2, ammonium sulfate 0.1, magnesium sulfate 0.05, dipotassium hydrogen phosphate 0.1, calcium chloride 0.02, yeast 0.03, and beef extract 0.03.
2. Determination of crude enzyme Activity
2.1 p-nitrophenol method (p NP method)
mu.L of 1mM rho NPR/rho NPG + 300. mu.L of 50 mM pH6 citric acid buffer solution, incubating for 2 min, adding 100. mu.L of crude enzyme solution, reacting at 50 deg.C for 5min, adding 500. mu.L of 1M Na2CO3The reaction was stopped and the absorbance was measured at 405 nm.
The standard curve is shown in figure 3, and the enzyme activity measurement result is shown in figure 4, wherein the measurement result of the rhamnosidase enzyme activity is 0.0068U/mL, and the measurement result of the glucosidase enzyme activity is 0.117U/mL.
Example 3: serratia marcescensSerratia marcescensAnalysis of naringin dihydrochalcone catalyzed and hydrolyzed by naringin produced by C10
The fermentation broth prepared in example 2 was centrifuged at 10000 r/min at 4 ℃ for 10min, the supernatant was discarded, 10mL of 50 mM PBS buffer pH8.0 was added, the centrifugation was carried out at 10000 r/min at 4 ℃ for 5min, the operation was repeated twice, the supernatant was discarded, and finally PBS buffer was added at 1:10 (g: mL) for ultrasonication. The ultrasonic crushing condition is 200W, the crushing time is 2 s, the stopping time is 2 s, and the time is 10 min. Taking 2mL of 0.25mg/mL naringin dihydrochalcone, 1mL of 50 mM of citric acid buffer solution with pH6 and 1mL of crude enzyme solution, carrying out water bath reaction at 50 ℃ for 30min, and then inactivating the mixture in water bath at 100 ℃ for 10 min.
HPLC conditions of high performance liquid chromatography detection are that a chromatographic column is a Wondassil C18 analytical column (250 mm multiplied by 4.6 mm, 5 mu m), a mobile phase A is 0.1% formic acid water, a mobile phase B is acetonitrile, the speed is 0.9mL/min, the ultraviolet detection wavelength is 283nm, the column temperature is 35 ℃, the sample injection volume is 20 mu L, and the elution program is 0-1 min 5% A; 1-5 min, 75% A; 60% A for 5-15 min; 50% A for 15-25 min; 85% A for 25-30 min, and the speed is 1.00 mL/min. The conversion rate = naringin dihydrochalcone content before reaction (mg)/naringin dihydrochalcone total amount after reaction (mg) × 100% was calculated. The results are shown in FIG. 5.
The result shows that the efficiency of naringin catalytic hydrolysis of naringin dihydrochalcone by the naringin enzyme produced by the serratia marcescens C10 is as high as 89.3%.
Example 4: serratia marcescensSerratia marcescensC10 naringinase production condition optimization
3.1 optimization of culture conditions
The growth and metabolism of bacteria mainly comprise three stages, wherein the first stage is the stage of bacteria rapid growth and propagation, the second stage is the stage of bacteria nearly stopping growth and propagation and then performing target protein expression, and the last stage is the stage of bacterial thallus autolysis cracking due to environmental deterioration. The most important time for the strains we screened to express the enzyme protein of interest is in the second stage of bacterial growth, so we optimize the culture conditions in the second stage and hope to obtain the optimal enzyme production conditions for fermentation to obtain a higher proportion of enzyme protein. The influence of factors such as the seed amount, the fermentation temperature, the fermentation time and the like on the expression form of the target protein is discussed respectively.
3.1.1 optimization of inoculum size
Inoculating the seed solution into an expression culture medium according to the proportion of 0.5, 1, 1.5 and 2%, culturing at 180 rpm and 30 ℃ overnight (about 24 h), centrifugally collecting thalli at 4 ℃ after the culture is finished, suspending the thalli by precooled PBS, centrifugally crushing the thalli by ultrasonic waves at 4 ℃ and 12000 rpm for 15min, collecting supernatant, and re-suspending precipitates by PBS. The content of the soluble target protein is measured respectively.
3.1.2 optimization of fermentation temperature
Inoculating 1% seed solution into expression culture medium, culturing at 180 rpm at 20, 25, 30, 35, and 40 deg.C overnight (about 24 h), centrifuging at 4 deg.C to collect thallus, suspending thallus with precooled PBS, ultrasonically crushing, centrifuging at 4 deg.C and 12000 rpm for 15min, collecting supernatant, and resuspending the precipitate with PBS. The content of the soluble target protein is measured respectively.
3.1.3 optimization of fermentation time
Inoculating 1% seed solution into an expression culture medium, culturing at 180 rpm and 30 ℃ for 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 and 30 hours respectively, centrifuging at 4 ℃ after the culture is finished, collecting thalli, suspending the thalli by precooled PBS, crushing by ultrasonic waves, centrifuging at 4 ℃ and 12000 rpm for 15min, collecting supernatant, and suspending the precipitate by PBS. The content of the soluble target protein is measured respectively.
The experimental results show that the inoculation amount is 1 percent; the fermentation time is 24 h; the best effect of enzyme production by fermentation is achieved when the fermentation temperature is 30 ℃.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Sequence listing
<110> university of southern China's science
Shandong Benyue Biotechnology Ltd
<120> Serratia marcescens and application thereof in naringinase production
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gtaatgtctg ggaaactgcc tgatggaggg ggataactac tggaaacggt agctaatacc 120
gcataacgtc gcaagaccaa agagggggac cttcgggcct cttgccatca gatgtgccca 180
gatgggatta gctagtaggt ggggtaatgg ctcacctagg cgacgatccc tagctggtct 240
gagaggatga ccagccacac tggaactgag acacggtcca gactcctacg ggaggcagca 300
gtggggaata ttgcacaatg ggcgcaagcc tgatgcagcc atgccgcgtg tgtgaagaag 360
gccttcgggt tgtaaagcac tttcagcgag gaggaaggtg gtgagcttaa tacgctcatc 420
aattgacgtt actcgcagaa gaagcaccgg ctaactccgt gccagcagcc gcggtaatac 480
ggagggtgca agcgttaatc ggaattactg ggcgtaaagc gcacgcaggc ggtttgttaa 540
gtcagatgtg aaatccccgg gctcaacctg ggaactgcat ttgaaactgg caagctagag 600
tctcgtagag gggggtagaa ttccaggtgt agcggtgaaa tgcgtagaga tctggaggaa 660
taccggtggc gaaggcggcc ccctggacga agactgacgc tcaggtgcga aagcgtgggg 720
agcaaacagg attagatacc ctggtagtcc acgctgtaaa cgatgtcgat ttggaggttg 780
tgcccttgag gcgtggcttc cggagctaac gcgttaaatc gaccgcctgg ggagtacggc 840
cgcaaggtta aaactcaaat gaattgacgg gggcccgcac aagcggtgga gcatgtggtt 900
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tggattggtg ccttcgggaa ctctgagaca ggtgctgcat ggctgtcgtc agctcgtgtt 1020
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Claims (9)
1. A serratia marcescens strain is characterized in that: designated as Serratia marcescens (Serratia marcescens) C10, deposited at No. 59 of the great college, Jie Lianlu 100, Calif., Guangdong province, in Guangzhou, Youzhou, 1/19, 2022, with the deposit number GDMCC No. 62223.
2. Use of the serratia marcescens C10 as claimed in claim 1 for the production of naringinase.
3. The use of serratia marcescens C10 for the production of naringinase according to claim 2 wherein: the naringinase consists of alpha-L-rhamnosidase and beta-D-glucosidase.
4. A method for producing naringinase by using the Serratia marcescens C10 as claimed in claim 1, wherein: the method comprises the following steps:
(1) inoculating serratia marcescens C10 to a solid culture medium for culture, and then inoculating to a liquid culture medium for culture to obtain a fermentation liquid;
(2) centrifugally separating the fermentation liquor, discarding the supernatant, resuspending the precipitate, and ultrasonically crushing to obtain a cell crushing liquid;
(3) separating and purifying the cell disruption solution to obtain naringinase.
5. The method of claim 4, wherein:
the solid culture medium inoculated in the step (1) is cultured for 1-3 days at the temperature of 28-32 ℃ and at the speed of 150-200 r/min;
the solid medium described in step (1) contains per 100 mL: 0.15-0.25 g of naringin, 0.05-0.15 g of ammonium sulfate, 0.01-0.1 g of magnesium sulfate, 0.05-0.15 g of dipotassium hydrogen phosphate, 0.01-0.03 g of calcium chloride, 0.02-0.04 g of yeast, 0.02-0.04 g of beef extract, 0.5-1.5 g of agar powder and the balance of water;
the conditions of inoculating the strain in the liquid culture medium and culturing in the step (1) are that the strain is cultured for 1-3 days at the temperature of 28-32 ℃ and at the speed of 150-200 r/min;
the liquid medium described in step (1) contains per 100 mL: 0.15-0.25 g of naringin, 0.05-0.15 g of ammonium sulfate, 0.01-0.1 g of magnesium sulfate, 0.05-0.15 g of dipotassium hydrogen phosphate, 0.01-0.03 g of calcium chloride, 0.02-0.04 g of yeast, 0.02-0.04 g of beef extract and the balance of water;
the inoculation amount of the inoculation in the liquid culture medium for culture in the step (1) is 1-2%;
the centrifugation condition in the step (2) is 8000-12000 r/min, and the centrifugation is carried out for 8-12 min at the temperature of 3-5 ℃;
the reagent used for resuspension in the step (2) is PBS buffer solution with pH8.0 +/-0.5, and the dosage is calculated according to the precipitation reagent =1g: 5-15 mL;
and (3) carrying out ultrasonic treatment in the step (2) under the condition of 150-250W, crushing for 2 s, stopping for 2 s, and carrying out time 10 min.
6. The method of claim 4, wherein:
the solid culture medium inoculated in the step (1) is cultured for 3 days at the temperature of 30 ℃ and at the speed of 180 r/min;
the solid medium described in step (1) contains per 100 mL: naringin 0.2 g, ammonium sulfate 0.1 g, magnesium sulfate 0.05 g, dipotassium hydrogen phosphate 0.1 g, calcium chloride 0.02 g, yeast 0.03 g, beef extract 0.03 g, agar powder 1g and water in balance;
the conditions of inoculating the strain in the liquid culture medium and culturing in the step (1) are 30 ℃ and 180r/min for 1 day;
the liquid medium described in step (1) contains per 100 mL: naringin 0.2 g, ammonium sulfate 0.1 g, magnesium sulfate 0.05 g, dipotassium hydrogen phosphate 0.1 g, calcium chloride 0.02 g, yeast 0.03 g, beef extract 0.03 g, and water in balance;
centrifuging for 10min at 4 ℃ under the conditions of 10000 r/min for the centrifuging in the step (2);
the reagent for resuspension in step (2) is PBS buffer solution with pH8.0 +/-0.5, and the dosage is calculated according to the precipitation reagent =1g: 10 mL;
and (3) the ultrasonic condition in the step (2) is 200W, crushing is carried out for 2 s, stopping for 2 s, and the time is 10 min.
7. A biological agent having naringinase activity, comprising: a cell containing the Serratia marcescens C10 according to claim 1.
8. Use of the serratia marcescens C10 as claimed in claim 1 or the biological agent with naringinase activity as claimed in claim 7 in the preparation of an agent for hydrolyzing naringin.
9. Use according to claim 8, characterized in that:
the application comprises the following steps: mixing naringin, cell disruption solution of serratia marcescens C10 and citric acid buffer solution, reacting at 45-55 ℃ for 25-35 min, and inactivating at 95-100 ℃ for 8-12 min;
the citric acid buffer solution is 45-55 mmol/L of citric acid buffer solution with the pH value of 6 +/-0.2;
the proportion of the naringin, the cell disruption solution of the serratia marcescens C10 and the citric acid buffer solution is 4-6 mg, 1mL and 1 mL.
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