1. A Rahnella (Rahnella sp.) HGS-393 strain for converting ginsenoside Rf into Rh1, the preservation number of the strain is as follows: CCTCC NO: m2021498, the 16S rDNA sequence of the strain is shown in SEQ ID NO. 1.

2. The use of the rahnella HGS-393 strain of claim 1, wherein: used for preparing ginsenoside Rh1 by fermentation and transformation.

Background

Ginseng (Panax ginseng) is a traditional Chinese medicinal material in China. Ginsenoside (Ginsenoside) is the main active medicinal component of ginseng, has a plurality of physiological and pharmacological activities of resisting tumor, resisting oxidation, protecting cardiovascular system and nervous system, and the like, and is widely applied to the development of medicaments and health care products.

Ginsenosides are classified into prototype ginsenosides and rare ginsenosides. The prototype ginsenoside refers to ginsenoside extracted from Ginseng radix, and comprises Rf, Ra, Rb1, etc. The rare ginsenoside refers to ginsenoside component with stronger bioactivity formed by metabolic conversion of prototype ginsenoside, and is low or free in Panax plant, including Rh1, Rg3, C-K, Rh2, etc.

Compared with prototype ginsenoside, the rare ginsenoside has more remarkable biological activity and pharmacological activity. The rare ginsenoside Rh1 has good effects on preventing and treating diseases such as Alzheimer disease, atherosclerosis, rheumatoid arthritis and the like, and can be widely applied to clinic as an anti-tumor auxiliary preparation.

The rare ginsenoside is generally obtained by converting a prototype ginsenoside by physical, chemical, or microbial methods. Physical and chemical methods are easy to destroy the aglycone structure in the conversion process, and have low conversion rate, violent reaction and more byproducts. The microbial conversion method has the advantages of high conversion efficiency, strong specificity, low energy consumption, no pollution and the like.

Disclosure of Invention

The invention solves the problems in the background technology and provides a Rahnella (Rahnella sp.) HGS-393 strain for converting ginsenoside Rf into Rh1, the strain can convert the ginsenoside Rf into the ginsenoside Rh1, and the conversion rate is high.

The inventor selects a novel strain which is named as HGS-393 and is a Laenheir (Rahnella sp.) strain, and the strain is preserved in China center for type culture Collection with the address: china, wuhan university; the zip code 430072 has a preservation date of 2021, 5 months and 7 days, and has a preservation number of: CCTCC NO: m2021498; the 16S rDNA sequence of the strain is shown in SEQ ID NO. 1.

The Rahnella strain HGS-393 can be used for preparing ginsenoside Rh1 by fermentation and transformation.

Compared with the prior art, the invention has the following advantages: the Rahnella HGS-393 strain provided by the invention can convert prototype ginsenoside Rf into rare ginsenoside Rh 1; the cost of the strain for transforming and producing the ginsenoside Rh1 is low, the reaction is fast, the transformation rate is high, and the transformation efficiency of transforming the prototype ginsenoside Rf fermented for 20 hours into the rare ginsenoside Rh1 is as high as 59.62%. During the reaction of producing ginsenoside Rh1 by transforming the strain, the target product is clear, the interference of other impurities is small, the ginsenoside Rh1 is easier to prepare, and the strain has obvious advantages in the process of producing Rh1 by mass fermentation.

Drawings

FIG. 1 is an HPLC detection chart of ginsenoside Rf and ginsenoside Rh1 in the fermentation broth before the fermentation conversion of Rahnella HGS-393 strain;

FIG. 2 is an HPLC detection chart of ginsenoside Rf and ginsenoside Rh1 in the fermentation liquid after fermentation and conversion of Rahnella strain HGS-393;

FIG. 3 is an LC-MS single-channel directional detection diagram of ginsenoside Rf in the fermentation broth after fermentation and transformation of Rahnella HGS-393 strain;

FIG. 4 is an LC-MS single-channel directional detection diagram of ginsenoside Rh1 in the fermentation liquid after fermentation and transformation of Rahnella HGS-393 strain.

Detailed Description

The present invention will be described in detail with reference to specific examples, but the scope of the present invention is not limited to the examples.

Isolation and culture of the Strain

In the embodiment, 0.5g of root tissue of 5-year-old fresh wild ginseng collected from Changbai mountain is surface-sterilized with 75% ethanol for 20min, washed with sterile water for 3-5 times, put into a mortar sterilized in advance for standby use, added with 5mL of sterile water, ground into a suspension, uniformly coated on an LA culture medium plate, inversely cultured in a 37 ℃ constant temperature incubator for 3-5 days, picked with a toothpick, transferred to a new LA plate, and repeatedly purified until a single colony is obtained.

The monoclonal bacteria are picked into a 1.5mL EP tube filled with 600 mu L LB liquid medium, shake-cultured for 1d at 37 ℃ and 190r/min, added with 50% glycerol with the same volume, mixed evenly and stored in an ultra-low temperature refrigerator at minus 80 ℃, and each strain is stored at least 3 parts.

Screening of strains

Inoculating 10 μ L of the preserved strain into 10mL LB liquid culture medium, performing activation culture at 37 deg.C for 24 hr by shaking at constant temperature of 180r/min, fermenting according to 5% transfer amount to 100mL LB liquid culture medium (liquid loading amount of 100mL/250mL), and shaking at 37 deg.C for 180r/min to OD₆₀₀When the concentration is 0.8, ginsenoside Rf is added to a final concentration of 30 mg/L. After 20h fermentation culture, collecting fermentation liquor.

And (3) carrying out HPLC detection on each strain fermentation liquor sample, and comparing the HPLC spectrograms and data of the standard ginsenoside Rf and Rh1 to determine whether the strains can directionally convert Rf into Rh 1. And calculating the transformation efficiency of the target strain to Rf according to the standard curve of the corresponding ginsenoside.

The ginseng endophyte capable of converting ginsenoside Rf into ginsenoside Rh1 is obtained by screening after the detection, and is named as HGS-393 strain. The conversion efficiency of the ginsenoside Rf which is fermented for 20 hours by the strain to the ginsenoside Rh1 is 59.62 percent, and the HPLC detection of a fermentation solution before fermentation conversion is shown in figure 1; HPLC detection of fermentation broth after fermentation conversion is shown in FIG. 2.

The applicant adopts LC-MS single-channel directional detection on the fermentation liquor after the fermentation and conversion of the HGS-393 strain, and the detection results are shown in figures 3 and 4, so that the HGS-393 strain is further determined to be capable of converting ginsenoside Rf into ginsenoside Rh 1.

Sequencing, sequence alignment and analysis

The strain is sent to a biotechnology limited company for sequencing, and the 16S rDNA sequence of the strain is shown as SEQ ID NO. 1. Sequencing results are subjected to Blast similarity analysis in a GenBank nucleic acid database, and are determined to be Laenna (Rahnella sp.) through Blast sequence alignment and evolutionary tree analysis.

Preservation of the strains:

the strain is preserved in China center for type culture Collection with the address: china, wuhan university; the zip code 430072 has a preservation date of 2021, 5 months and 7 days, and has a preservation number of: CCTCC NO: m2021498.

Sequence listing

<110> university of the south China nationality

<120> Laenna HGS-393 strain for converting ginsenoside Rf into Rh1 and application

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1436

<212> DNA

<213> genus Rahnella (Rahnella sp.)

<400> 1

ttggggcagc taccatgcag tcgagcggca gcggaaagta gcttgctact ttgccggcga 60

gcggcggacg ggtgagtaat gtctgggaaa ctgcctgatg gagggggata actactggaa 120

acggtagcta ataccgcatg acctcgaaag agcaaagtgg gggatcttcg gacctcacgc 180

catcggatgt gcccagatgg gattagctag taggtgaggt aatggctcac ctaggcgacg 240

atccctagct ggtctgagag gatgaccagc cacactggaa ctgagacacg gtccagactc 300

ctacgggagg cagcagtggg gaatattgca caatgggcgc aagcctgatg cagccatgcc 360

gcgtgtgtga agaaggcctt agggttgtaa agcactttca gcgaggagga aggcatcata 420

cttaatacgt gtggtgattg acgttactcg cagaagaagc accggctaac tccgtgccag 480

cagccgcggt aatacggagg gtgcaagcgt taatcggaat tactgggcgt aaagcgcacg 540

caggcggttt gttaagtcag atgtgaaatc cccgcgctta acgtgggaac tgcatttgaa 600

actggcaagc tagagtcttg tagagggggg tagaattcca ggtgtagcgg tgaaatgcgt 660

agagatctgg aggaataccg gtggcgaagg cggccccctg gacaaagact gacgctcagg 720

tgcgaaagcg tggggagcaa acaggattag ataccctggt agtccacgct gtaaacgatg 780

tcgacttgga ggttgtgccc ttgaggcgtg gcttccggag ctaacgcgtt aagtcgaccg 840

cctggggagt acggccgcaa ggttaaaact caaatgaatt gacgggggcc cgcacaagcg 900

gtggagcatg tggtttaatt cgatgcaacg cgaagaacct tacctactct tgacatccac 960

ggaattcgcc agagatggct tagtgccttc gggaaccgtg agacaggtgc tgcatggctg 1020

tcgtcagctc gtgttgtgaa atgttgggtt aagtcccgca acgagcgcaa cccttatcct 1080

ttgttgccag cgagtcatgt cgggaactca aaggagactg ccggtgataa accggaggaa 1140

ggtggggatg acgtcaagtc atcatggccc ttacgagtag ggctacacac gtgctacaat 1200

ggcatataca aagagaagcg aactcgcgag agcaagcgga cctcataaag tatgtcgtag 1260

tccggattgg agtctgcaac tcgactccat gaagtcggaa tcgctagtaa tcgtagatca 1320

gaatgctacg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca ccatgggagt 1380

gggttgcaaa agaagtaggt agcttaacct tcgggagggc gctaccactg gatcct 1436