NL2026395B1 - Method for deodorization of sargassum fusiforme - Google Patents

Abstract

The invention relates to the field of microorganisms and food processing, and specifically refers to a method for deodorization of Sargassum fusiforme by fermentation with Cordyceps militaris. The method of the present invention features simple process, low cost, good deodorization effect for Sargassum fusiforme, and easy realization of industrialized production.

Description

1 001627P-NL

METHOD FOR DEODORIZATION OF SARGASSUM FUSIFORME FIELD OF TECHNOLOGY

[0001] The invention relates to the field of microorganisms and food processing, and specifically refers to a method for deodorization of Sargassum fusiforme by fermentation with Cordvceps militaris.

BACKGROUND

[0002] Sargassum fusiforme has a unique flavor, is rich in nutrients such as polysaccharides, dietary fiber, vitamins and minerals, and has the effects of eliminating brain fatigue, enhancing immunity, delaying aging, lowering blood sugar, and lowering blood lipids (Zhongneng Xu, Zhaohui Wang, Li Sun. Research progress on the medicinal value of Sargassum fusiforme. Chinese Traditional and Herbal Drugs, 2000. 31(11). 876-878.). Sargassum fusiforme is a marine vegetable and traditional delicacy found in coastal areas of Japan, China, South Korea and North Korea. The Japanese are particularly fond of Sargassum Jfusiforme, and they have highly praised it as a longevity food since ancient times (Zhan Zhang, Jianguo Liu, Jidong Liu. Research review of Sargassum fusiforme. Marine Fisheries Research, 2002. 23(3): 67-74.).

[0003] Seaweed is nutritious and delicious, but has a strong fishy smell, which has seriously affected consumer acceptance and the development of the seaweed industry. At present, people have higher and higher requirements for food flavor. How to control the formation of fishy substances and remove them has attracted more and more attention of researchers. For the fishy smell of seaweed, the main fishy substance removal technologies include acid-base treatment, masking, steam deodorization, adsorption, microencapsulation, extraction, and microbial fermentation. CN101496562 discloses a method for deodorizing kelp, where the fishy smell of kelp is removed by hot blanching with sour plum soup and soaking in tea; CN101218974 discloses a method for treating Sargassum fusiforme with a chlorine dioxide bleaching solution for color protection and deodorization; CN102326799 discloses a multi-step method for deodorization of Sargassum fusiforme using a licorice solution and ozone water; CN104187878 discloses a deodorized kelp powder product and its

2 001627P-NL production method, where pickled kelp as a raw material is subjected to a two-stage treatment with a tea polyphenol solution and an alcohol of above 95% to prepare a deodorized kelp powder. Although traditional deodorizing methods such as acid-base treatment, masking, and adsorption have a certain deodorizing effect on seaweed, they may damage nutrients such as proteins to a certain extent, thereby affecting the nutritional value of seaweed. Microbial deodorizing is an environmentally friendly deodorizing method with good deodorizing effect, which not only improves the flavor of seaweed, but also can improve its biological activity (Xianyan Zhang, Wenyang Duan, Jiaxin Zou, Jidan Li, Hailong Yang*. Research progress in seaweed deodorization technology. Journal of Food Safety and Quality Inspection, 2017, 8(11): 4288-4293).

SUMMARY

[0004] An objective of the present invention is to provide a method for deodorization of Sargassum fusiforme by microbial fermentation.

[0005] In order to achieve the above-mentioned objective, the technical solution of the present invention adopts Cordvceps militaris deposited in the China General Microbiological Culture Collection Center with Deposit No. CGMCC5.856. The relevant properties of the bacterial strain can be referred to the list of strains of the China General Microbiological Culture Collection Center (CCGMC) and will not be detailed in the present invention.

[0006] (1) inocula culture: the Cordvceps militaris 1s inoculated to a soybean sprout culture medium, and subjected to rotatory submerged fermentation at a temperature of 23-25°C and a rotation speed of 140-180 r/min for 5-7 days; the culture medium comprises 200 g/L soybean sprouts (boiling for 30 min, extracting juice), 20-30 g/L glucose, 1-3 g/L peptone, and 3-5 g/L corn flour;

[0007] (2) fermentation: dried Sargassum fusiforme is crushed and passed through a 60-100 mesh sieve; 100 g of the Sargassum fusiforme powder is weighed and put into a fermentation container, 80-100 mL of distilled water is added thereto, the mixture is well stirred, sterilized at 121°C for 30 min; after cooling, it is inoculated with a 10-15% (v/w) Cordyceps militaris seed fermentation broth, well stirred and allowed to stand still at 25-28°C

3 001627P-NL to ferment for 12-15 days; the fermented Sargassum fusiforme has no fishy smell at all.

[0008] The positive and beneficial effects of the present invention are:

[0009] The method of the present invention features simple process, low cost, good deodorization effect for Sargassum fusiforme, and easy realization of industrialized production.

BRIEF DESCRIPTION OF THE DRAWINGS

[0010] In order to explain the embodiments of the present invention or the technical solutions in the prior art more clearly, the following will briefly introduce the drawings that need to be used in the description of the embodiments of the present invention or the prior art. Obviously, the drawings in the following description are only are some embodiments of the present invention. For those of ordinary skill in the art, other drawings obtained based on these drawings without creative labor still belongs to the scope of the present invention.

[0011] Figure 1 is a GC-MS total ion chromatogram of volatile components in unfermented Sargassum fusiforme;

[0012] Figure 2 is a GC-MS total ion chromatogram of volatile components of Sargassum fusiforme fermented with Cordyceps militaris.

DESCRIPTION OF THE EMBODIMENTS

[0013] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below with reference to the attached drawings.

[0014] In the following examples, the experimental methods without specific conditions are selected according to conventional methods and conditions, or according to the product specification.

[0015] Example 1 GC-MS analysis of volatile components of Sargassum fusiforme

[0016] (1) Sample processing

[0017] First, the SPME extraction fiber head was aged at 250°C at the GC-MS inlet until there was no impurity peak. 5.0 g of a sample was accurately weighed and put into a sample bottle, then 100 uL of an internal standard (4-methyl-2-pentanol solution,

4 001627P-NL concentration 4.6 mg/ml) was added thereto, and the cap of the bottle was screwed on for sealing. The temperature of the solid phase microextraction device was set to 80°C, the sample bottle was placed in the extraction device to preheat for 15 min; the SPME extraction head was inserted through the bottle cap into the headspace of the sample so that it was about

1.5 cm above the upper surface of the sample, and the fiber head was pushed out to conduct headspace extraction for 60 min; the fiber head was withdrawn, and the extraction head was pulled out from the sample bottle; then the extraction head was inserted into the GC-MS injection port, the fiber was taken out for desorption at 250°C for 5 min, and then the sample was injected for analysis.

[0018] (2) Instruments and measurement conditions

[0019] Agilent gas chromatography-mass spectrometer GC 6890-MS5975.

[0020] Chromatographic conditions: chromatographic column, HP-INNOWAX (60.0 m x 250 um, 0.25 pum); initial temperature, 35°C, held for 5min, then increased to 230°C at a rate of 5°C/min and held for 8 min; gasification chamber temperature, 25°C; transfer line temperature, 220°C; carrier gas, He; carrier gas flow rate, 1.0 mL/min; splitless injection.

[0021] Mass spectrometry conditions: electron impact ion source (EI), electron energy 70 eV, ion source temperature 230°C, quadrupole 150°C; scanning mode: Scan; scanning mass range: 30-500 u.

[0022] (3) Data processing

[0023] The MS database NISTII and retention time were used for qualitative analysis of the detected components. The relative quantification method of internal standard was used to quantify each component.

[0024] Example 2

[0025] Cordyceps militaris CGMCC5.856 was inoculated into a soybean sprout inocula culture medium for rotatory submerged fermentation at a temperature of 25°C and a rotation speed of 150 r/min for 5 days. The dried Sargassum fusiforme was crushed and passed through an 80-mesh sieve. 100 g of the Sargassum fusiforme powder was weighed and put into a fermentation container, 80-100 mL distilled water was added thereto, the mixture was well stirred, and sterilized at 121°C for 30 min. After cooling, it was inoculated with a

001627P-NL 15% (v/w) Cordvceps militaris seed fermentation broth, well stirred, and allowed to stand still to ferment at 28°C for 12 days. The volatile components of Sargassum fusiforme before and after fermentation were tested according to the method of Example 1. The total ion chromatograms of the volatile components of unfermented Sargassum fusiforme and 5 Sargassum fusiforme fermented with Cordvceps militaris are shown in Figs | and 2. It can be seen from the figures that fermentation with Cordvceps militaris significantly reduces the volatile components of Sargassum fusiforme. The volatile components of unfermented Sargassum fusiforme are shown in Table 1, and the volatile components of Sargassum Jusiforme fermented with Cordyceps militaris are shown in Table 2. Fermentation with Cordvceps militaris significantly reduces the number and content of volatile components of Sargassum fusiforme and makes Sargassum fusiforme free of fishy smell.

[0026] Example 3

[0027] Cordyceps militaris CGMCC5.856 was inoculated into a soybean sprout inocula culture medium for rotatory submerged fermentation at a temperature of 25°C and a rotation speed of 150 r/min for 7 days. The dried Sargassum fusiforme was crushed and passed through a 60-mesh sieve. 100 g of the Sargassum fusiforme powder was weighed and put into a fermentation container, 80-100 mL distilled water was added thereto, the mixture was well stirred, and sterilized at 121°C for 30 min. After cooling, it was inoculated with a 15% (v/w) Cordyceps militaris seed fermentation broth, well stirred, and allowed to stand still to ferment at 28°C for 15 days. After the fermentation, the volatile components of the Sargassum fusiforme fermented with Cordyceps militaris were analyzed by the GC-MS method, and a similar result as in Example 2 was achieved. The Sargassum fusiforme fermented with Cordyceps militaris had no fishy smell.

[0028] Example 4

[0029] Cordyceps militaris CGMCC5.856 was inoculated into a soybean sprout inocula culture medium for rotatory submerged fermentation at a temperature of 24°C and a rotation speed of 180 r/min for 7 days. The dried Sargassum fusiforme was crushed and passed through a 100-mesh sieve. 100 g of the Sargassum fusiforme powder was weighed and put into a fermentation container, 80-100 mL distilled water was added thereto, the mixture was well stirred, and sterilized at 121°C for 30 min. After cooling, it was inoculated with a

6 001627P-NL 15% (v/w) Cordvceps militaris seed fermentation broth, well stirred, and allowed to stand still to ferment at 28°C for 10 days. After the fermentation, the volatile components of the Sargassum fusiforme fermented with Cordyceps militaris were analyzed by the GC-MS method, and a similar result as in Example 2 was achieved. The Sargassum fusiforme fermented with Cordyceps militaris had no fishy smell.

[0030] The above-disclosed are only preferred embodiments of the present invention. Of course, the scope of protection of the present invention cannot be limited thereto. Therefore, equivalent changes made according to the claims of the present invention still fall within the scope of the present invention.

[0031] Table 1 Volatile components of unfermented Sargassum fusiforme

[0032] Table 2 Volatile components of Sargassum fusiforme fermented with Cordyceps militaris

[0033] Table 1 . . Molecular Content

1.142 Acetic acid, methyl ester 74.037 000079-20-9 1.10 Ethyl Acetate 88.052 000141-78-6 15.00

3.502 3-Furanol, tetrahvdro- 88.052 000453-20-3 0.46 Butanal, 3-methyl- 86.073 000390-86-3

9.979 Hexane, 3,3-dimethyl- 114.141 000563-16-6 0.55

12.081 Nonane, 5-butyl- 184.219 017312-63-9

12.918 Trichloroacetic acid, 2-0ctyl ester 274.029 344883-74-5 0.64

14.068 2-Propanone, 1-cyclopropyl- 98.073 004160-75-2 0.28

15.068 Undecane, 2-methyl- 170.203 007045-71-8 1-Penten-3-ol 86.073 000616-25-1

7 001627P-NL

16.683 Dodecane 170.203 000112-40-3 3.13 cthvlcarbazole

8 001627P-NL

27.055 Dodecyl acrylate 240.209 002156-97-0 0.92

27.395 140.157 019150-21-1 0.64

27.849 2.5-Furandicarboxaldehyde 124.016 000823-82-5

27.995 3,5-Octadien-2-one. (E.E)- 124.089 030086-02-3

28.157 2-Furancarboxaldchyde, 110.037 000620-02-0 0.46 5-methyl-

28.351 | Ethanone, 1-(1-cyclohexen-1-vl)- 124.089 000932-66-1

28.562 | 6-Methyl-3.5-heptadiene-2-one 124.089 001604-28-0

28.837 Cyclohexanol. 2,6-dimethyl- 128.12 005337-72-4

28.886 2-Octen-1-ol, (E)- 128.12 018409-17-1 0.18 Ethanone, J 0.18 2 > oy C Ol. .

29.172 | | .methyl 1-oyolopenten-Li)- 124.089 003168-90-9 1-Cyclohexene-1-carboxaldehyde, a 331

29.318 36. trimethyl. 152.12 000432-25-7 1,3-Cvclohexadiene-1-carboxalde Ce 2 85 (C 5 2 6. .

29.939 ede. 2.6.6.4r methyl. 150.104 000116-26-7

30.096 2-Furanmethanol 98.037 000098-00-0 0.74

30.264 Butanoic acid, 3-methyl- 102.068 000503-74-2

1.3-Cyelooctadiene 108.094 001700-10-3 : 1,4-Hexadiene, A Tend 2 0.18 . ; 5 A34. .

30.642 23.4 totramothal- 138.141 051504-54-2

31.085 2,6,6-Trimethyl-2-cyelohexene- 1, 152.084 001125-21-9 0.46 4-dione 2-Thiapentane, 0.83

31.436 | 4-(9-borabicyclo[3.3.1]non-9-ylo 226.156 1000162-69-3 xy)-

31.96 Oxime-, methoxy-phenyl-_ 151.063 1000222-86-6

32.057 9-Borabicyclo[3.3.1 Jnonane. 208.2 1000160-72-1 2.48 9-(2-pentyloxy)- Naphthalene, 1.01

32.295 L2-diydron]. 16 teimethyl- 172.125 030364-38-6

32.581 | Chloromcetie acid: Z-ethylhexst 206.107 005345-58-4

32.711 (E)-4-Oxohex-2-enal 112.052 1000374-04-2

33.672 Imidazole-5-pentanoic acid 168.09 1000126-21-3

34.018 -Methyl-2-methylenecyclohexan 110.11 002808-75-5

34.504 3,9-Undecadien-2-0ne, 194.167 000689-67-8 0.46 6,10-dimethyl-

34.834 Naphthalene. 2-methyl- 142.078 000091-57-6

34.947 | Butyric acid, 4-pentadecyl ester 298.287 1000280-57-4

35.087 Benzyl alcohol 108.058 000100-31-6 0.46 Benzene, oy 2.12

5.2 : C 4-19. 2.

9 001627P-NL EE: DE | owns |

37.589 4-(2,2,6-trimethy1-7-oxabicyclo[4. 208.146 023267-57-4

1.0]hept-1-y1)- | won [rien] vos | wo | OE

44.611 5,6,7,7a+tetrahydro-4,4,7a-trimeth 180.115 015356-74-8 yL Table 2

001627P-NL A Sprro[5.5]undec-2-ene, J ; 2.48

23.208 B,7,7-trimethyl- 1 1-methylene-, (-)- 204.188 018431-82-8

23.624 3-Cyclohexen-1-one. 138.104 000471-01-2 0.64

3.5.5-trimethyl- quinolinamine

24.391 Benzene, 1.2.3 4-tetramethyl- 134.11 000488-23-3

24.564 128.12 003391-86-4 0.74

24.688 60.021 000064-19-7 60.63 25569 Acetic acid, trichloro-, heptyl ester | 260.014 065611-31-6 Lo Benzaldehvde, 0.4 0.74 24: | Berzalachiye 56114-69- :

26.244 2.5-bis[(trimethylsilyl)oxv]- 282.111 056114-69-3

26.93 Benzaldehyde 106.042 000100-52-7 Bicyclo[3.1.1]hept-2-ene, 1.20

28.254 6-dimethyl-6-(4-methyl-3-pentenyl | 204.188 017699-05-7 )-

28.691 138.104 000078-59-1 0.46 Benzoic acid, 0.28

29.707 2 4-bis[(trimethylsilyloxy]-, 370.145 010586-16-0 trimethylsilyl ester

30.269 3-Methyl-hexanoic acid 130.099 1000365-65-4

31.96 Oxime-, methoxy-phenyl- 151.063 1000222-86-6

32.057 Benzene, 1.3-dimethoxy- 138.068 000151-10-0 42 861 71013. 16-Hexaoxacyelooctadee 264.157 017455-13-9

Claims (1)

11 001627P-EN Conclusions 1. A method of deodorizing Sargassum fusiforme by fermentation with Cordyceps militaris, the method being implemented using C'ordyceps militaris purchased from China General Microbiological Culture Collection Center with deposit number CGMCC5.856. The method consists of the following steps: (1) seed culture: the Cordvceps militaris is inoculated on a soybean germ culture medium and subjected to rotary submerged fermentation at a temperature of 23-25°C and a rotational speed of 140-180 rpm for 5-7 days; the culture medium comprises 200 g/l soybean sprouts (cooking for 30 minutes, extracting juice), 20-30 g/l glucose, 1-3 g/l peptone and 3-5 µl maize flour, (2) fermentation: dried Sargassum fusiforme is crushed and passed through a 60-100 mesh screen; 100 g of Sargassum fusiforme powder is weighed and put into a fermentation vessel; 80-100 ml of distilled water are added thereto: the mixture is stirred well and sterilized at 121°C for 30 minutes; after cooling, it is inoculated with a 10-15% (v/w) Cordyceps militaris seed fermentation broth; stir well and then stand still for 12-15 days at 25-28°C to ferment; the fermented Sargassum fusiforme has no fishy smell at all.