Dried Fish Fermentation Process and Starter Culture Development Technology

Abstract

The present invention provides a dried fish fermentation process and a starter culture development technology. A mixed starter culture including Bifidobacterium species, Lactobacillus acidophilus, Lactobacillus casei, Streptococcus thermophilus and Bifidobacterium lactis is prepared for the mixed fermentation of a dried fish; and salt content in the dried fish is controlled to control a proportional relation of flora in the mixed starter culture and to optimize the fermentation process, thereby preparing more fresh and delicious, fishy-free, more soft and delicate dried fish. Therefore, the dried fish is more popular with consumers.

Claims

1. A mixed starter culture of a dried fish, comprising Bifidobacterium species, Lactobacillus acidophilus, Lactobacillus casei, Streptococcus thermophilus and Bifidobacterium lactis. wherein in the mixed starter culture, Bifidobacterium species, Lactobacillus acidophilus, Lactobacillus casei, Streptococcus thermophilus and Bifidobacterium lactis have a volume ratio of 1:1:1:2; and the mixed starter culture has a concentration of bacterium of 10.sup.8-10.sup.9 cfu/mL.

2. (canceled)

3. A method for preparing a dried fish, comprising the following steps: preparing a dried fish: taking and scaling off freshwater fish, then dissecting and killing, and pickling the fish with salt which is 15%-20% weight of the fish body for 12 h after being cleaned; then cleaning the pickled fish with running water, draining off and drying the fish to control a water content to 35%; rinsing the dried fish with fresh water: subjecting the dried fish to desalting treatment for 1-2 h, and then baking the dried fish at a condition of 40° C., and after baking, the weight of the dried fish being 1.1-1.2 times of the original weight; and inoculating with the mixed starter culture as claimed in claim 1 for fermentation, wherein the mixed starter culture has an inoculum size of 10-20 mL/100 g dried fish, the inoculation way is spray or rolling and rubbing, and sealed fermentation is performed for 24 h at a fermentation temperature of 20° C.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0035] FIG. 1 is a gas chromatography-mass spectrometry diagram of a dried fish prepared by mixed fermentation in Example 6;

[0036] FIG. 2 is a gas chromatography-mass spectrometry diagram of non-fermented dried fish in Example 6.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0037] The present invention will be further described by reference to the examples. It should be indicated that the following examples are aimed at understanding the present invention, but not construed as limiting the present invention. Reagents in this example are known products purchased from the market available.

Example 1 Preparation of the Mixed Fermented Dried Fish

[0038] The preparation method of the mixed fermented dried fish provided in this example is as follows:

[0039] (1) Preparation of Dried Fish:

[0040] a freshwater fish (grass carp) was taken, scaled off, dissected and killed, pickled with salt which was 20% weight of the fish body for 12 h after being cleaned; then the pickled fish was cleaned with running water to remove residues and surface stains, and then drained off and dried at 40° C. to control a water content to 35% around; the dried fish was cut into pieces having a side length of about 5 cm, and salt content was about 10%.

[0041] (2) Rinsing the Dried Fish with Fresh Water:

[0042] the dried fish was taken and rinsed with fresh water for 1 h, and water was exchanged twice, where, the ratio of raw material to water was 1:15. the rinsed dried fish was drained off to remove surface water, and then put to a constant temperature drying oven at 40° C. for drying until 1.1-1.2 times of the raw dried fish before rinsing with fresh water; salt content in the dried fish after being rinsed and dried was about 2%;

[0043] (3) Inoculation with the Mixed Starter Culture for Fermentation:

[0044] a. Preparation of the Mixed Starter Culture:

[0045] Bifidobacterium species powder (purchased from China Center of Industrial Culture Collection (CICC) No. 21711), Lactobacillus acidophilus powder (purchased from CICC No. 20244), Lactobacillus casei (purchased from CICC No. 20241), Streptococcus thermophilus (purchased from Mufan Biotechnology Company No. MF-001031) and Bifidobacterium lactis powder (purchased from Mufan Biotechnology Company No. MF-001040) were respectively taken and inoculated onto MRS culture media (purchased from Qingdao Hopebio, type HB0384-1) with an inoculum size of 5%, and then cultured over the night at 30° C.;

[0046] the activated bacterial solutions of Bifidobacterium species, Lactobacillus acidophilus, Lactobacillus casei, Streptococcus thermophilus and Bifidobacterium lactis were then respectively taken and inoculated onto domestication culture media (50% MRS culture media and 50% fish broth) for domestication culture for 24 h at 30° C. with an inoculum size of 10% to respectively obtain the bacterial solutions of Bifidobacterium species, Lactobacillus acidophilus, Lactobacillus casei, Streptococcus thermophilus and Bifidobacterium lactis having a concentration of 10.sup.8-10.sup.9 cfu/m L;

[0047] the bacterial solutions of Bifidobacterium species, Lactobacillus acidophilus, Lactobacillus casei, Streptococcus thermophilus and Bifidobacterium lactis were mixed according to a volume ratio of 1:1:1:2 to prepare into a mixed starter culture;

[0048] b. Mixed Fermentation

[0049] the mixed starter culture was inoculated to the rinsed dried fish with an inoculum size of 15 mL/100 g dried fish via spray or rolling and rubbing, and stirred to be mixed well; then the mixture was seal fermented for 24 h at 20° C. the fermented dried fish was put to a constant temperature drying oven for drying at 40° C. until the original weight of the dried fish before rinsing with fresh water; then, the fermented dried fish product was obtained.

Example 2 Influences of Different Fermented Starters on the Prepared Dried Fish

[0050] In this example, the dried fish was prepared by the method provided in Example 1; during fermentation, different starters (the starters used are shown in Table 1; starters were mixed according to the same volume in the combination of bacterial solution) were inoculated to the dried fish with an inoculum size of 15 mL/100 g dried fish and concentration of 10.sup.8-10.sup.9 cfu/mL. The contents of flavor substances, 5-hexyldihydro-2(3H)-furanone (peach flavor), 3-heptyldihydro-5-methyl-2(3H)-furanone (green onion/garlic flavor), nonanal (waxy and fat flavor), hexadecanal (flower and wax flavor), and delicious substances, glutamic acid and aspartic acid in the prepared dried fish as well as the fishy smell and taste were detected. The content of flavor substances was detected by gas chromatography-mass spectrography, and the content of delicious substances was reflected by the content of free amino acids; fishy smell and taste were subjected to comprehensive evaluation after being tasted by 30 volunteers, thereby exploring the influences of different fermented starters on the flavor substances, delicious substances and flavor of the prepared dried fish. The results are shown in Table 1.

TABLE-US-00001 TABLE 1 Influences of different fermented starters on the prepared dried fish 3-heptyldihydro- 5-hexyldihydro- 5-methyl-2(3H)- Nonanal Hexadecanal Glutamic Aspartic Fishy Starter 2(3H)-furanone furanone (%) (%) acid (%) acid (%) smell Taste Bifidobacterium 0.71 1.58 0.65 1.77 8.5 6.7 Slightly Soft species fishy Lactobacillus 0.83 1.82 0.72 1.83 9.1 7.5 Slightly Soft acidophilus fishy Lactobacillus 0.94 1.92 0.83 2.32 8.7 7.2 No fishy Slightly casei smell hard Streptococcus 1.01 1.75 0.66 1.64 8.5 8.9 Slightly Slightly thermophilus fishy hard Bifidobacterium 1.52 2.46 0.88 3.08 9.8 9.5 No fishy Soft lactis smell Bifidobacterium 1.54 2.51 0.87 3.09 9.7 9.8 No fishy Soft lactis + smell Bifidobacterium species Bifidobacterium 1.53 2.55 0.85 3.11 9.5 10.7 No fishy Soft lactis + smell Lactobacillus acidophilus Bifidobacterium 1.84 2.65 0.91 3.17 10.4 12.3 No fishy Slightly lactis + smell soft Lactobacillus casei Bifidobacterium 1.65 2.63 0.89 3.15 9.8 11.3 No fishy Slightly lactis + smell soft Streptococcus thermophilus Bifidobacterium 1.93 2.96 0.87 3.30 10.9 12.5 No fishy More soft lactis + smell and waxy Bifidobacterium species + Lactobacillus acidophilus Bifidobacterium 2.01 3.24 0.98 3.58 11.8 14.4 No fishy Soft lactis + smell Lactobacillus casei + Streptococcus thermophilus Bifidobacterium 1.94 3.05 0.89 3.21 11.1 12.8 No fishy More soft lactis + smell and waxy Bifidobacterium species + Lactobacillus acidophilus + Lactobacillus casei Bifidobacterium 2.25 3.87 1.01 4.03 12.6 16.8 No fishy Very lactis + smell delicious Bifidobacterium and waxy species + Lactobacillus acidophilus + Lactobacillus casei + Streptococcus thermophilus

[0051] It can be seen from Table 1 that the fermentation with a single bacterium may slightly increase the flavor substances and delicious substances in dried fish, but the effect is not obvious; the result is that the dried fish tastes not enough delicious and waxy, and sometimes slightly hard, and even the fishy smell of the dried fish may not be completely removed; the effect of Bifidobacterium lactis in improving flavor substances and delicious substances in dried fish is slightly better than other bacteria, and the effect of removing fishy smell is better slightly.

[0052] It can be seen that during the combined fermentation of several bacteria, the combination of the five bacteria according to the same volume for mixed fermentation has an excellent effect of improving the quality of dried fish, obviously better than the mixed fermentation effects of the two, three and four bacteria. The reason may be because the five bacteria create a synergistic effect during mixed fermentation to promote the production of more flavor and delicious substances in the pickled fish. The contents of 5-hexyldihydro-2(3H)-furanone, 3-heptyldihydro-5-methyl-2(3H)-furanone, nonanal, hexadecanal, glutamic acid and aspartic acid are up to 2.25%, 3.87%, 1.01%, 4.03%, 12.6% and 16.8%. The dried fish tastes very good, delicious and waxy.

Example 3 Influences of the Content of Bifidobacterium lactis in the Mixed Starter Culture on the Prepared Dried Fish

[0053] In this example, the dried fish was prepared by the method provided in Example 1; during mixed fermentation, the proportional relation of Bifidobacterium lactis in the mixed bacterial solution was changed (see details in Table 2); then, the mixed bacterial solution was respectively inoculated to dried fish for mixed fermentation with an inoculum size of 15 mL/100 g dried fish and concentration of 10.sup.8-10.sup.9 cfu/mL. The contents of flavor substances, 5-hexyldihydro-2(3H)-furanone (peach flavor), 3-heptyldihydro-5-methyl-2(3H)-furanone (green onion/garlic flavor), nonanal (wax and fat flavor), hexadecanal (flower and wax flavor), and delicious substances, glutamic acid and aspartic acid in the prepared dried fish as well as the fishy smell and taste were detected. The detection method is the same as that in Example 2. The results are shown in Table 2.

TABLE-US-00002 TABLE 2 Influences of the proportional relation of Bifidobacterium lactis in the mixed starter culture on the prepared dried fish Bifidobacterium species:Lactobacillus acidophilus:Lactobacillus casei:Streptococcus 3-heptyldihydro- thermophilus:Bifidobacterium 5-hexyldihydro- 5-methyl-2(3H)- Nonanal Hexadecanal Glutamic Aspartic Fishy lactis 2(3H)-furanone furanone (%) (%) acid (%) acid (%) smell Taste 1:1:1:1:1 2.25 3.87 1.01 4.03 12.6 16.8 No fishy Very smell delicious and waxy 1:1:1:1:1.5 2.28 3.98 1.01 4.14 13.1 17.0 No fishy Very smell delicious and waxy 1:1:1:1:2 2.34 4.03 1.02 4.27 13.5 17.7 No fishy Very smell delicious, waxy and delicate 1:1:1:1:3 2.01 2.94 0.95 3.24 10.7 15.5 No fishy Too soft smell

[0054] It can be seen from Table 2 that the proportional relation of Bifidobacterium lactis in the mixed bacterial solution has greater influences on the quality of dried fish; when the ratio of Bifidobacterium species: Lactobacillus acidophilus: Lactobacillus casei: Streptococcus thermophilus: Bifidobacterium lactis is up to 1:1:1:1:2, the content of flavor and delicious substances in the pickled fish is up to the maximum; and the contents of 5-hexyldihydro-2(3H)-furanone, 3-heptyldihydro-5-methyl-2(3H)-furanone, nonanal, hexadecanal, glutamic acid and aspartic acid are respectively up to 2.34%, 4.03%, 1.02%, 4.27%, 13.5% and 17.7%. The dried fish has an excellent taste, free of fishy smell, very delicious, waxy and delicate. When the ratio of Bifidobacterium lactis is further increased, the content of various flavor and delicious substances significantly decreases; moreover, the prepared dried fish tastes too soft and not very good. Therefore, the ratio of Bifidobacterium species, Lactobacillus acidophilus, Lactobacillus casei, Streptococcus thermophilus and Bifidobacterium lactis is controlled to 1:1:1:1:2, thus preparing dried fish with better quality.

Example 4 Influences of Salt Content on the Prepared Dried Fish after being Rinsed with Fresh Water

[0055] In this example, the dried fish was prepared by the method provided in Example 1; the salt content in the dried fish was controlled by controlling the rinsing time. It is proved by experiments that the dried fish has a salt content of 6% after being rinsed with fresh water for 30 min, has a salt content of 2% after being rinsed with fresh water for 60 min, and has a salt content of 0.1% after being rinsed with fresh water for 90 min; and then the dried fish was subjected to mixed fermentation respectively. The contents of flavor substances, 5-hexyldihydro-2(3H)-furanone (peach flavor), 3-heptyldihydro-5-methyl-2(3H)-furanone (green onion/garlic flavor), nonanal (waxy and fat flavor), hexadecanal (flower and wax flavor), and delicious substances, glutamic acid and aspartic acid in the prepared dried fish as well as the fishy smell and taste were detected. The detection method is the same as that in Example 2. The results are shown in Table 3.

TABLE-US-00003 TABLE 3 Influences of salt content on the prepared dried fish after being rinsed with fresh water 3-heptyldihydro- Rinsing 5-hexyldihydro- 5-methyl-2(3H)- Nonanal Hexadecanal Glutamic Aspartic Fishy time 2(3H)-furanone furanone (%) (%) acid (%) acid (%) smell Taste 30 min 0.79 1.34 0.85 1.97 8.1 10.7 No fishy Slightly smell hard 60 min 2.34 4.03 1.02 4.27 13.5 17.7 No fishy Very smell delicious, waxy and delicate 90 min 2.17 3.02 0.98 3.75 12.4 15.6 No fishy Too soft smell

[0056] It can be seen from Table 3 that the rinsing time has greater influences on the quality of the dried fish. The reason is that the salt content in the dried fish varies from the rinsing time; and the salt content in the dried fish will directly influence the effect of the mixed fermentation. When the rinsing time is short (30 min), the salt content in the dried fish is higher, and if the mixed fermentation is performed at this time, the mixed fermentation may be inhibited, leading to not enough flavor and delicious substances in the dried fish. When the rinsing time is too long (90 min), the salt content in the dried fish is very low, and if the mixed fermentation is performed at this time, the quality of the prepared dried fish declines. It can be seen that the rinsing time is controlled to keep a certain salt content in the dried fish, which is also an important condition to prepare mixed fermented dried fish with high quality; that is, the rinsing time may provide more suitable fermentation conditions for the mixed fermentation, thus promoting the increase of flavor and delicious substances in the dried fish, and improving taste. Therefore, the rinsing time needs to be controlled to 60 min.

Example 5 Influences of Inoculum Size of the Mixed Starter Culture on the Prepared Dried Fish

[0057] In this example, the dried fish was prepared by the method provided in Example 1; the inoculum size of the mixed starter culture was respectively selected from 5, 10, 15, 20 and 25 mL/100 g (as shown in Table 4. The contents of flavor substances, 5-hexyldihydro-2(3H)-furanone (peach flavor), 3-heptyldihydro-5-methyl-2(3H)-furanone (green onion/garlic flavor), nonanal (wax and fat flavor), hexadecanal (flower and wax flavor), and delicious substances, glutamic acid and aspartic acid in the prepared dried fish as well as the fishy smell and taste were detected. The detection method is the same as that in Example 2. The results are shown in Table 4.

TABLE-US-00004 TABLE 4 Influences of inoculum size of the mixed starter culture on the prepared dried fish Inoculum size 3-heptyldihydro- (mL/100 g 5-hexyldihydro- 5-methyl-2(3H)- Nonanal Hexadecanal Glutamic Aspartic Fishy dried fish) 2(3H)-furanone furanone (%) (%) acid (%) acid (%) smell Taste 5 1.35 1.89 0.89 2.57 10.7 13.6 Slightly Slightly fishy hard 10 2.24 3.97 0.99 4.01 12.9 16.1 No fishy Very smell delicious, waxy and delicate 15 2.35 4.04 1.02 4.28 13.5 17.6 No fishy Very smell delicious, waxy and delicate 20 2.19 3.95 0.95 4.03 12.7 16.5 No fishy Very smell delicious, waxy and delicate 25 2.01 3.58 0.89 3.95 12.0 15.7 No fishy Too soft smell

[0058] It can be seen from Table 4 that a proper inoculum size is selected to obtain better quality of dried fish. When the inoculum size of the mixed starter culture is 10-20 mL/100 g dried fish, the prepared dried fish has a higher content of flavor and delicious substances, tastes delicious, waxy and delicate, free of fishy smell. The optimal inoculum size is 15 mL/100 g dried fish; and at this time, the prepared dried fish has the highest content of flavor and delicious substances, and tastes excellent.

Example 6 Comparison Between the Dried Fish Prepared in this Present Invention and the Dried Fish without Mixed Fermentation

[0059] In this example, the dried fish was prepared by the method provided in Example 1; the dried fish without mixed fermentation (the dried fish prepared in the step one of Example 1) served as a control group for comparison. The content of volatile substances in the two kinds of dried fish was detected by gas chromatography-mass spectrometry; and the content of delicious substances was detected (reflected by the content of free amino acids); color, smell, taste and state of the two groups of dried fish were scored. There are 10 scores in total, more than 8 represents excellent quality (white and light yellow, aromatic and delicate flavor, good taste, slightly sweet residual taste, soft and waxy and moderate meat quality); more than 6 represents good quality, and less than 6 represents poor quality (brown, obvious fishy smell, spicy flavor, bitter taste of residual taste, too hard or too soft meat). The gas chromatography-mass spectrometry results are shown in FIGS. 1-2 (FIG. 1 is a detection diagram of the dried fish prepared by mixed fermentation provided in Example 1; FIG. 2 is a detection diagram of the dried fish in the control group) and Tables 5-6 (Table 5 shows the detection results of the dried fish prepared by mixed fermentation provided in Example 1; Table 6 shows the detection results of the dried fish in the control group). Detection results of the delicious substances, and score results of the color, smell, taste and state are shown in Table 7.

TABLE-US-00005 TABLE 5 Chemical contents of volatile substances in the dried fish prepared by mixed fermentation Relative Aroma percentage Name of volatile substances characteristics (%) Ketone 5 5-hexyldihydro-2(3H)-furanone Peach flavor 2.34 7.67 3-heptyldihydro-5-methyl-2(3H)-furanone Green onion/ 4.03 garlic flavor 2-nonanone Fruity and sweet 0.39 2-hendecanone Wax and fat 0.59 flavor 2-Tridecanone Milky and 0.32 coconut flavor Benzene 1 3-allyl-6-metoxybenzene 2.7 2.7 Acid 2 n-palmitic acid 3.65 4.5 Myristic acid 0.85 Ester 2 Ethyl cetylate Weak waxy and 1.32 2.11 cream flavor N-butyl-isooctyl phthalate Orange flavor 0.79 Alkene 1 Styrene Slightly sweet 0.69 0.69 Alcohol 5 1-octene-3-ol Mushroom flavor 4.46 7.43 2-phenethyl alcohol Rose flavor 0.62 3-octanol 1.01 1-octanol Strong grease 0.59 smell and rose flavor 2-octene-1-ol 0.75 Aldehyde 3 Nonanal Waxy and fat 1.01 6.37 flavor Hexadecanal Weak flowery 4.27 and wax flavor N-undecylic aldehyde Fat and flowery 1.09 flavor Others 1 Dimethyl trisulfide 2.27 2.27 Alkane 9 Dodecane 1.69 11.96 N-decane 0.69 Nonadecane 0.79 Octamethylcyclotetrasiloxane 0.75 N-hexadecane 0.72 Dodecamethylcyclohexasiloxane 0.72 Cetylcyclooctasiloxane 1.59 Hexamethylcyclotrisiloxane 3.19 Tetradecamethylcycloheptasiloxane 1.82

TABLE-US-00006 TABLE 6 Chemical contents of volatile substances of the dried fish in the control group Relative Aroma percentage Name of volatile substances characteristics (%) Ketone 3 3-octanone Fruit aroma 1.15 3.6 5-hexyldihydro-2(3H)-furanone Peach flavor 0.71 3-heptyldihydro-5-methyl-2(3H)-furanone Green onion/ 1.74 garlic flavor Phenols 2 Eugenol Clove flavor 14.26 14.85 Phenol Special odor, and 0.59 highly-diluted solution is sweet Benzene 2 P-xylene Aromatic flavor 0.67 1.01 Ethylbenzene 0.34 Acid 3 n-palmitic acid 1.86 2.44 Myristic acid 0.44 cis-octadecenoic acid 0.14 Ester 4 Dibutyl phthalate Aromatic flavor 0.6 2.49 Ethyl myristate 1.09 Ethyl cetylate Orange flavor 0.58 Methyl palmitate 0.22 Alkene 2 Styrene 0.72 1.37 Caryophyllene Spicy, wood and 0.65 orange flavor Alcohol 4 1-octene-3-ol Mushroom 1.61 2.71 flavor 2-phenethyl alcohol Rose flavor 0.52 Hendecanol 0.42 3-octanol 0.16 Aldehyde 3 Nonanal Waxy and fat 0.63 2.77 flavor Hexadecanal Weak flowery 1.66 and wax flavor Cis-9-hexadecenal 0.48 Others 2 Methoxybenzene oxime 3.5 6.98 Benzpyrole Highly-diluted 3.48 solution is fragrant Alkane 14 Dodecane 2.14 19.63 N-decane 1.11 Heptadecane 3.94 Octamethylcyclotetrasiloxane 0.37 N-tetradecane 1.46 N-hexadecane 0.49 Dodecamethylcyclohexasiloxane 0.29 Dodecamethylcyclohexasiloxane 1.15 Cetylcyclooctasiloxane 2.07 8-propoxy cedulane 0.35 Hexamethylcyclotrisiloxane 1.48 Octadecmethylcyclononasiloxane 2.88 Dimethyldiacetoxysilane 0.48 Hexadecmethyloctasiloxane 1.42

TABLE-US-00007 TABLE 7 Detection results of delicious substances in the two groups of dried fish Glutamic Aspartic Group acid (%) acid (%) Color Smell Taste State Mixed fermented 13.5 17.6 9 9 9 9 dried fish Control group 8.3 6.5 6 5 5 6

[0060] It can be seen from FIGS. 1-2 and Tables 5-6 that the dried fish prepared by mixed fermentation provided herein produces a large number of volatile flavor substances to achieve an aromatic flavor and excellent taste based on the original dried fish (control group). It can be seen from Table 7 that the dried fish prepared by mixed fermentation provided herein has obviously improved delicious substances, yellow and bright color, aromatic and fresh flavor, slightly sweet residual taste, soft and waxy meat; and the quality is promoted greatly. The dried fish will be well received by consumers.

[0061] The present invention is disclosed above, but the present invention is not limited thereto. Any person skilled in the art may make various alterations and modifications within the spirit and scope of the present invention. Therefore, the protection scope of the present invention shall be subjected to the scope defined in the claims appended.