KR101975105B1 - Kazachstania servazzii strain with high sugar degrading activity and gas producing activity isolated from apple pickle solution - Google Patents

Abstract

The present invention relates to a method for producing a sweet potato starch from Kazakhstania The present invention relates to a strain KM 2017 (KCCM 11989P), which comprises a strain of Kazakhstania serovar servazzii ) KM 2017 strain has excellent sucrose or lactose decomposing ability and shows excellent whey efficacy. Also, it has excellent gas generating ability and can be useful as a fermentation seed for baking and food material development.

Description

[0001] The present invention relates to a strain of Kazakhstania servazzii having high glucose degradation activity and gas generating activity isolated from apple pickles,

(KCCM11989P), which is excellent in the sugar resolution and gas production ability separated from apple pickles in Kazakhstania servazzii KM 2017 strain.

Yeast is a useful microorganism that is used as an important host in the production of industrial enzymes and various physiological functional materials and biotechnology in recent years.

Kazachstania servazzii ) were first introduced to Saccharomyces servazzii ), but the Kazachstania genus belonging to the Saccharomycetaceae family was first classified as a single species by Kazachstania viticola , a genus isolated from the grapes fermented in Kazakhstan in 1971, The strain name was changed from Saccharomyces servazzii to Kazachstania servazzii . Kazachstania servazzii strains were isolated from kimchi and cheese in Korea and abroad, but glucose utilization was not reported to be active.

Lactose is a disaccharide in which galactose and glucose are combined. It is also called lactose and lactose. It is mainly produced in the mother's milk of a mammal. Its structure is β-D-galactosyl- (1,4) -D-glucose . There are two types, α and β, depending on the type. Lactose is also a component of mammalian milk, and it is mainly used as a raw material for formulating formula for childcare and as a medical nutrition. Lactose is known to be used only by some yeast.

Korean Patent Publication No. 10-2017-0038555 (Apr.

It is an object of the present invention to provide a process for producing a sweet potato starch from Kazakhstania servazzii ) KM 2017 strain (KCCM11989P).

In order to accomplish the above object, the present invention provides a method for producing a saccharide- servazzii ) KM 2017 strain (KCCM11989P).

The present invention also provides a strain of Kazachstania servazzii KM 2017 (KCCM11989P) which is excellent in whey efficacy.

In addition, the present invention relates to the above-mentioned Kazakhstania serbia servazzii < / RTI > KM < RTI ID = 0.0 > 2017 < / RTI >

The present invention relates to a method for producing a sweet potato starch from Kazakhstania The present invention relates to a strain KM 2017 (KCCM 11989P), which comprises a strain of Kazakhstania serovar servazzii ) KM 2017 strain has excellent sucrose or lactose decomposing ability and shows excellent whey efficacy. Also, it has excellent gas generating ability and can be useful as a fermentation seed for baking and food material development.

Fig. 1 shows a process for separating yeast from apple pickles.
Fig. 2 shows the result of phylogenetic tree neighboring joining phylogram analysis of yeast isolated from apple pickles. apple: Kazachstania servazzii KM 2017 strain.
Figures 3 and 4 show the DNA sequence of yeast isolated from apple pickles.
FIG. 5 is a cross- servazzii) it shows a process diagram for the experimental gas producing ability of strain 2017 KM.
FIG. 6 is a graphical representation of the < RTI ID = 0.0 & The results of pH and acidity of the KM 2017 strain for gas production experiments are shown. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
FIG. 7 is a graphical representation of the < RTI ID = 0.0 & The results of the sugar content of the KM 2017 strain are shown. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Fig. 8 is a graphical representation of the < RTI ID = 0.0 > The results of measurement of viable cell count for the gas production ability of KM 2017 strain are shown. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Fig. 9 is a graph showing the results obtained by measuring the content of the baker's yeast Saccharomyces cerevisiae and Kazachstania < RTI ID = 0.0 > servazzii ) KM 2017 strain. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Figure 10 is a graphical representation of the < RTI ID = 0.0 > servazzii) it shows a process diagram for the gas producing ability test (PDB medium) of strain 2017 KM.
Fig. 11 is a graphical representation of the < RTI ID = 0.0 & The results of pH and acidity for the gas production experiment (PDB medium) of KM 2017 strain are shown. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Figure 12 is a cross- servazzii) shows the sugar content results for gas producing ability test (PDB medium) of strain 2017 KM. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Fig. 13 is a graphical representation of the < RTI ID = 0.0 > servazzii) it shows the number of viable cells results of measurements of experimental gas producing ability (PDB medium) of strain 2017 KM. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Figure 14 is a graphical representation of the < RTI ID = 0.0 > servazzii) shows the evaluation results for the product gas to a gas producing ability test (PDB medium) of strain 2017 KM. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Figure 15 is a graphical representation of the < RTI ID = 0.0 > servazzii) gas producing ability of the test KM 2017 strain (represents a process diagram for the YPD medium).
Figure 16 is a graphical representation of the < RTI ID = 0.0 > The results of pH and acidity of the KM 2017 strain (YPD medium) are shown. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Figure 17 is a graphical representation of the < RTI ID = 0.0 > servazzii) gas producing ability of the test KM 2017 strain (indicating the sugar content results in YPD medium). ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Figure 18 is a graphical representation of the < RTI ID = 0.0 > servazzii) gas producing ability of the test KM 2017 strain (represents the number of viable cells measured results for YPD medium). ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Figure 19 is a graphical representation of the < RTI ID = 0.0 > servazzii) gas producing ability of the test KM 2017 strain (shows a gas producing ability results for YPD medium). ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
FIG. 20 is a graphical representation of the < RTI ID = 0.0 & lt ; RTI ID = 0.0 > KM < / RTI > 2017 strain.
Figure 21 is a graphical representation of the < RTI ID = 0.0 > The pH and acidity results of the lactose degrading ability of KM 2017 strain are shown. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Figure 22 is a graphical representation of the < RTI ID = 0.0 > The present invention relates to a method for producing lactose, ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Fig. 23 is a graphical representation of the < RTI ID = servazzii ) KM 2017 strains. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Figure 24 is a graphical representation of the < RTI ID = 0.0 > servazzii ) KM 2017 strain of the present invention. ¹⁾ S. cerevisiae , ²⁾ K. servazzii .
Figure 25 is a graphical representation of the < RTI ID = 0.0 > servazzii ) KM 2017 strain according to the lactose concentration.
Figure 26 is a graphical representation of the < RTI ID = 0.0 > The pH and acidity of the KM 2017 strain according to the lactose concentration are shown. ¹⁾ lactose 0 mL, ²⁾ lactose 13 mL, and ³⁾ lactose 25 mL.
Figure 27 is a graphical representation of the < RTI ID = 0.0 > The results of the sugar content of the KM 2017 strain according to the lactose concentration are shown. ¹⁾ lactose 0 mL, ²⁾ lactose 13 mL, and ³⁾ lactose 25 mL.
28 is a cross- servazzii ) KM 2017 strain by the lactose concentration. ¹⁾ lactose 0 mL, ²⁾ lactose 13 mL, and ³⁾ lactose 25 mL.
FIG. 29 is a graphical representation of the < RTI ID = 0.0 > The results of gas production for the resolution according to the lactose concentration of KM 2017 strain are shown. ¹⁾ lactose 0 mL, ²⁾ lactose 13 mL, and ³⁾ lactose 25 mL.
Figure 30 is a graphical representation of the < RTI ID = 0.0 > servazzii ) KM 2017 strain using lactose according to whey concentration.
Figure 31 is a graph of The results of pH and acidity of the lactose application ability of KM 2017 strain according to whey concentration are shown. ^{1) No} dilution (undiluted solution), ²⁾ × 2 dilution.
Figure 32 is a graphical representation of the < RTI ID = 0.0 > servazzii ) KM 2017 strain according to whey concentration. ^{1) No} dilution (undiluted solution), ²⁾ × 2 dilution.
Figure 33 is a graphical representation of the < RTI ID = 0.0 > servazzii ) KM 2017 strain by lactic acid concentration. ^{1) No} dilution (undiluted solution), ²⁾ × 2 dilution.
Figure 34 is a graphical representation of the < RTI ID = 0.0 > servazzii ) KM 2017 by the whey concentration. ^{1) No} dilution (undiluted solution), ²⁾ × 2 dilution.

The present invention provides a strain of Kazakhstania servazzii KM 2017, which is entrusted with KCCM11989P and is excellent in sugar resolution and gas production ability .

Specifically, the saccharide may be sucrose or lactose, but is not limited thereto.

Specifically, the strain may be isolated from apple pickles, but is not limited thereto.

In addition, the present invention relates to a method for producing whey protein, which is entrusted with KCCM 11989P, servazzii ) KM 2017 strains.

The present invention also provides a whey fermentation method comprising culturing a Kazakhstania servazzii KM 2017 strain deposited with KCCM 11989P .

Specifically, the culture may be incubated at 25 to 30 DEG C for 1 to 5 days, but is not limited thereto.

The strain Kazachstania servazzii KM2017 used in the examples of the present invention was deposited as KCCM 11989P in Korean Microorganism Preservation Center.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> Apples From pickles  Yeast strain isolation

1. Experimental Method

10 μl of 10 ⁴ , 10 ⁵ , and 10 ⁶ times of diluted apple juice was applied to the MRS agar plate and cultured in a 30 ° C incubator for 24 to 48 hours. colony forming unit (CFU / mL). Subsequently, subculture was performed to identify microorganisms through ITS-1 and ITS-4 analysis methods (FIG. 1).

2. Experimental results

The Phylogenetic Tree is to identify similarities and differences in various biological species or other individuals, inferred evolutionary relationships of phylogeny, or physical or genetic characteristics. The nucleotide sequences were analyzed by Neighbor Joining, Maximum Likelihood, and Maximum Parsimony analysis using the phylogenetic tree . The homology was compared using the NCBI BLAST program. As a result, it was found that the homozygotes belonging to Kazacstania servazzii (Figs. 2 to 4).

< Example  2> Kazakhstania Serbazi ( Kazachstania servazzii ) Confirmation of gas production ability of KM 2017 strain

1. Experimental Method

As a result of the identification of the microorganisms isolated from apple pickles, Kazachstnia servazzii resulted in the following results. After adding 0.5 g of sucrose to 10 mL of MRS broth, the yeast saccharomyces cerevisiae S. cerevisiae and K. servazzii were inoculated 1%, and the culture was incubated for 3 days to compare pH, acidity, sugar content, viable cell count and gas production ability (FIG. 5).

2. Experimental results

(1) pH and acidity

Kazachstania servazzii) KM results of the pH and acidity for a gas producing ability of a test strain 2017 as Fig. The yeast tended to decrease pH while producing acid when fermented, and began to fermentation at pH 0 6.22 for S. cerevisiae inoculated samples. After the fermentation, the fermentation was slightly decreased on the first day of fermentation. On the other hand, in the case of samples inoculated with K. servazzii , the 0th day of fermentation was S. cerevisiae And the pH value was the same. However, the pH value of S. cerevisiae decreased to 5.82 on the first day of fermentation. And then decreased to pH 5.02 at the end of fermentation. In the case of acidity, both strains tended to increase with fermentation time.

(2) sugar content

Kazachstania servazzii) was changed is shown in Figure 7 of the sugar content of the gas producing ability of the test KM 2017 strain. The sugar content of early S. cerevisiae and K. servazzii were 5.8 ° Brix and 5.9 ° Brix, respectively. Afterwards, S. cerevisiae Inoculated samples were reduced to 4.2 ° Brix on the first day of fermentation, whereas 3 ° Brix was observed on samples inoculated with K. servazzi . After that, the sugar content of the fermented S. cerevisiae was 1.2 ° Brix and that of K. servazzi was 1 ° Brix.

(3) Measurement of viable cell count

Kazachstania The results of measurement of viable cell count for the gas production ability of KM 2017 strain are shown in FIG. The number of viable cells inoculated with S. cerevisiae started at 1.53 × 10 ⁶ CFU / mL on day 0 of fermentation. Followed by 5.41 × 10 ⁸ CFU / mL on the first day of fermentation. On the other hand, the K. servazzii inoculated samples showed 1.01 × 10 ⁹ CFU / mL. After three primary fermentation S. cerevisiae, K. All samples inoculated with servazzii showed a tendency to decrease in viable cell count. Thereafter, the viable counts of S. cerevisiae on the 5th day of fermentation were 2.83 × 10 ⁷ CFU / mL and 5.85 × 10 ⁷ CFU / mL for the samples inoculated with K. servazzii .

(4) Gas production experiment

Kazachstania The results of the gas production experiment of KM 2017 strain are shown in FIG. In this experiment, fermentation was carried out on the basis of 30 hours. In the case of S. cerevisiae , gas was produced at 1.2 hours in height and 2.6 cm in 12 hours. After 24 hours of fermentation, 7.2 cm of gas was produced. In contrast, in the sample inoculated with servazzii K. After 6 hours 3.5cm gas it is produced as a gas 10 cm after 18 hours fermentation S. cerevisiae And the gas generating ability was superior.

< Example  3> Kazakhstania Serbazi ( Kazachstania servazzii ) Confirmation of gas production ability of strain KM 2017 (PDB medium)

1. Experimental Method

Identification of microorganisms isolated from apple pickles Results came out with Kazachstania servazzii . The experiment was conducted by changing the nutrients. Experiments were carried out using Potato Dextrose Broth instead of conventional MRS broth. It is also referred to as crystalline glucose, and when the starch is sufficiently decomposed with acid, crystals are precipitated and separated from the glycosylated liquid to make crystalline glucose. Crystalline glucose has a sweet sweet taste and has almost the same sweetness as sucrose in crystals. The sweetness of the solution is about 60% of sucrose. The solubility in 1 L potato starch medium was 24 g. To make 250 g, 250 g of distilled water was added to 6 g of potato starch medium and sterilized at 121 ° C for 15 minutes. Each strain was then inoculated with 1% (FIG. 10 and Table 1).

A ¹⁾ B ²⁾ Potato dexrose broth 6 g 6 g Distilled water 250 g 250 g Yeast S. cerevisiae
2.5 mL (1%) K. servazzii
2.5 mL (1%)

¹⁾ S. cerevisiae , ²⁾ K. servazzii .

2. Experimental results

(1) pH and acidity

Kazachstania servazzii) KM results of the pH and acidity for a gas producing ability of a test strain 2017 as Fig. For the S. cerevisiae inoculated samples, the pH at 0 day of fermentation was 6.16. As the alcohol fermentation progressed, the pH was decreased and the fermentation showed a pH of 5.44 on the third day of fermentation. K. servazzii , The samples were inoculated with pH 6.18 at day 0 of fermentation. Then, S. cerevisiae It was confirmed that the pH was decreased to a larger extent. The pH of the fermentation 5 days later was recorded as 5.21. In the case of acidity, the samples inoculated with S. cerevisiae and K. servazzii tended to increase with fermentation.

(2) sugar content

Kazachstania servazzii) Results of sugar content for the gas producing ability of a test strain KM 2017 shown in Fig. S. cerevisiae and K. servazzii showed 5.7 ° Brix and 5.9 ° Brix, respectively. The first day of fermentation, S. cerevisiae showed a 4.3 ° Brix difference on the first day of fermentation. Thereafter, the sugar content was decreased to 2.5 ° Brix on the third day of fermentation. In the case of K. servazzii , the fermentation showed 3.2 ° Brix on the first day of fermentation and 2.1 ° Brix on the second day of fermentation. Thereafter, the fermentation showed a 1.4 ° Brix on the third day of fermentation.

(3) Number of living cells

Kazachstania servazzii) equal to the sweetness of the results of 13 experiments on the gas producing ability of strain 2017 KM. In the case of S. cerevisiae , the number of live cells was 2.51 × 10 ⁶ CFU / mL on day 0 of fermentation. Thereafter, the number of bacteria tended to increase to 5.13 × 10 ³ CFU / mL on the first day of fermentation. From the second day of fermentation, the number of bacteria decreased to 3.51 × 10 ⁷ CFU / mL on the third day of fermentation. For the samples inoculated with K. cerevisiae , the fermentation was expressed as 3.38 × 10 ⁶ CFU / mL on day 0 of fermentation. Thereafter, the number of bacteria in the first day of fermentation tended to increase and showed 2.80 × 10 ³ CFU / mL on the second day of fermentation. And then maintained at 10 ⁷ CFU / mL on the third day of fermentation.

(4) Gas production experiment

Kazachstania The results of the gas production experiment of KM 2017 strain are shown in FIG. In the case of S. cerevisiae inoculated, the gas was produced at 0.2 cm in height for 12 hours and at 2.6 cm in height for 18 hours after fermentation. For the samples inoculated with K. servazzii , the gas production of 12 hours of fermentation was recorded as 0.6 cm and the gas production ability of 6.2 cm was confirmed after 24 hours of fermentation.

< Example  4> Kazakhstania Serbazi ( Kazachstania servazzii ) Confirmation of gas production ability of strain KM 2017 (YPD medium)

1. Experimental Method

In this experiment, the experiment was conducted using YPD medium as a nutrient medium for culturing fungi and yeast. In contrast to other experiments, the former MRS and PDB media have been supplemented with sugars, but this experiment has no sugars. Then, the gas production ability of YPD medium was measured based on 20% glucose. 5 g of yeast extract and 10 g of peptone are added, followed by 25% (5%) of 20% glucose. After sterilization at 121 DEG C for 15 minutes, 4 mL of 1% strain was inoculated and cultured for 3 days (FIG. 15 and Table 2).

A ¹⁾ B ²⁾ yeast extract 5 g 5 g Peptone 10 g 10 g 20% glucose 25 mL 25 mL Yeast S. cerevisiae
4 mL (1%) K. servazzii
4 mL (1%)

¹⁾ S. cerevisiae , ²⁾ K. servazzii .

2. Experimental results

(1) pH and acidity

Kazachstania servazzii) KM results of the pH and acidity for a gas producing ability of a test strain 2017 as Fig. S. cerevisiae and K. servazzii were fermented at pH 6.06 and 6.10, respectively. In the case of K. servazzii inoculated on the 1st day after fermentation, it was confirmed that the pH decreased more than S. cerevisiae . Thereafter, each sample was decreased to pH 5.32 for samples inoculated with S. cerevisiae on the third day of fermentation, and 5.22 for samples inoculated with K. servazzii . In the case of acidity, both samples tended to increase with decreasing pH.

(2) sugar content

Kazachstania The sugar content of the KM 2017 strain is shown in FIG. 17. In the case of S. cerevisiae , the fermentation showed a day 0 difference of 6.4 ° Brix and decreased to 3.5 ° Brix on day 2 of fermentation. On the other hand, in the case of K. servazzi i, the fermentation showed a fermentation of 6.3 ° Brix on day 0 and a 2.9 ° Brix on day 2 of fermentation. Thereafter, the sugar content of each sample on the third day of fermentation decreased to 2.5 ° Brix and 1.1 ° Brix.

(3) Number of living cells

Kazachstania The results are shown in Fig. 18 for the viability of the KM 2017 strain. In the case of S. cerevisiae , fermentation showed 2.68 × 10 ⁶ CFU / mL on the 0th day of fermentation, and the bacterial strain tended to increase by 5.04 × 10 ³ CFU / mL on day 1 of fermentation. The number of viable cells in the second day of fermentation showed a tendency to decrease and was shown as 2.73 × 10 ⁷ CFU / mL on the third day of fermentation. K. servazzii The inoculated samples showed 2.87 × 10 ⁶ CFU / mL on day 0 of fermentation and showed 6.81 × 10 ³ CFU / mL on day 1 of fermentation. Thereafter, the viable cell count tended to decrease. And then expressed as 6.10 × 10 ⁷ CFU / mL on the third day of fermentation.

(4) Gas production experiment

Kazachstania The results of the gas production experiment of KM 2017 strain are shown in FIG. In the case of S. cerevisiae , 2.1 cm gas production occurred after 12 hours of fermentation. After 24 hours of fermentation, gas was produced at a height of 6.8 cm. On the other hand, for the samples inoculated with K. servazzii, 2.6 cm gas was generated after 6 hours of fermentation and 10 cm gas was produced after 24 hours of fermentation.

< Example  5> Kazakhstania Serbazi ( Kazachstania servazzii ) Confirmation of lactose utilization ability of strain KM 2017

1. Experimental Method

In this experiment, the lactose degradation test of K. servazzii , which was confirmed to be excellent in gas production, was carried out using YPD medium as a nutrient medium for the cultivation of fungi and yeast. Lactose is referred to as lactose or lactose and is the molecular formula C ₁₂ H ₂₂ O ₁₁ and has the structure of? -D-galactosyl- (1,4) -D-glucose. Lactose, as the name implies, is found in mammalian milk, especially in colostrum, which contains 6.7% in milk and 4.5% in milk. In breast milk, there are also more than 10 bovine sugars, all of which contain lactose as the skeleton. There have also been several reports that were found in plant seeds, although none were found in other than milk. There are two kinds of α and β type. α-Lactose has a slightly sweet taste. 1g is dissolved in 5mℓ of water and 2.6mℓ of hot water, but does not dissolve in alcohol and does not dissolve in chloroform or ether. β-lactose is formed when crystals are concentrated in a solution at a temperature of 93.5 ° C. or higher, which is more sweet than α-lactose. It is hydrolyzed by the action of dilute acid or enzyme (lactase) to become D-glucose and D-galactose, and lactic acid becomes lactic acid under action of lactic acid bacteria.

In this experiment, 5 g of yeast extract powder was added, 10 g of peptone was added, and 25 mL of 20% lactose was added in the same manner as in the previous YPD gas experiment. After sterilization, the cells were cultured for 5 days (FIG. 20 and Table 3).

A ¹⁾ B ²⁾ yeast extract 5 g 5 g Peptone 10 g 10 g 20% lactose 25 mL 25 mL Yeast S. cerevisiae
2.5 mL (1%) K. servazzii
2.5 mL (1%)

¹⁾ S. cerevisiae , ²⁾ K. servazzii .

2. Experimental results

(1) pH and acidity

Kazachstania The pH and acidity in the experiment of gas production and lactose decomposition of KM 2017 strain are shown in FIG. In the case of S. cerevisiae inoculated samples, pH and acidity did not change after fermentation. On the other hand, K. servazzii In the case of the sample inoculated with the bacteria, there was no change in pH until the first day of fermentation. However, fermentation showed a tendency to decrease slightly at pH 6.02 on the third day of fermentation and decreased to pH 5.83 on the 5th day of fermentation . The acidity of the samples inoculated with K servazzii did not change significantly until the first day of fermentation, whereas the acidity tended to increase slightly on the third day of fermentation.

(2) sugar content

Kazachstania The change in sugar content in the experiment of gas production and lactose decomposition of KM 2017 strain is shown in FIG. In the case of S. cerevisiae , the sugar content was 6.7 ° Brix even after fermentation, indicating that the sugar content did not decrease. On the other hand, the samples inoculated with K. servazzii tended to decrease slightly at 6 ° Brix on the third day of fermentation and decreased to 4.1 ° Brix on the fifth day of fermentation.

(3) Number of living cells

Kazachstania The change in the number of viable cells in the experiment of gas production and lactose decomposition of KM 2017 strain is shown in FIG. The samples inoculated with S. cerevisiae I smoked with a 10-fold dilution method, but no bacteria were found . K. servazzii The fermented samples showed 3.26 × 10 ⁵ CFU / mL on day 0 of fermentation and 7.4 × 10 ⁷ CFU / mL on day 1 of fermentation. From the second day of fermentation, viable cell counts were maintained. Fermentation Day 3 K. servazzii The viable cell count of the sample inoculated with the bacteria was 7.5 × 10 ⁷ CFU / mL.

(4) Gas production experiment

Kazachstania The results of the gas production in the experiment of gas production and lactose decomposition of KM 2017 strain are shown in FIG. In the case of S. cerevisiae inoculated, it was confirmed that no gas was produced even when the fermentation proceeded. On the other hand, for the samples inoculated with K. servazzii, gas was produced 1.2 cm high after 36 hours of fermentation and slightly increased to 2.5 cm after 48 hours of fermentation. It was confirmed that gas was produced up to 6.2 cm after 60 hours of fermentation.

< Example  6> Kazakhstania Serbazi ( Kazachstania servazzii ) Confirmation of the resolution of KM 2017 strain by lactose concentration

1. Experimental Method

(Lactose) was dissolved in YPD medium, which is a nutrient medium of fungi and yeast. As a result, 0 mL of lactose, 13 mL of lactose, and 25 mL of 0% (lactose) , 2.5%, 5%) was added to the test solution. As in the previous experiment, the lactose content was differentiated using a YPD medium containing no sugar (FIG. 25 and Table 4).

A B C Yeast extract 5 g 5 g 5 g Peptone 10 g 10 g 10 g 20% Lactose 0 mL 13 mL 25 mL D.W 25 mL 10 mL 0 mL Yeast K. servazzii
0.4 mL K. servazzii
0.4 mL K. servazzii
0.4 mL

2. Experimental results

(1) pH and acidity

Kazachstania The pH and acidity in the experiment of gas production and lactose decomposition of KM 2017 strain are shown in FIG. In the case of 20% lactose - free samples, the pH was 6.09 on day 0 of fermentation. For the sample containing 20 mL of lactose 13 mL, the fermentation showed a slight decrease with pH 6.02 on the third day of fermentation. The fermentation efficiency of yeast was lower than that of sucrose and glucose, but lactose was used. When 25 mL of 20% lactose was added, it was confirmed that the pH was decreased more than 13 mL of 20% glucose at pH 6.06 on the first day of fermentation, pH 5.84 on the third day of fermentation and pH 5.51 on the fifth day of fermentation. In the case of acidity, the acidity tended to increase as the fermentation proceeded, except for the sample without lactose.

(2) sugar content

Kazachstania The experiment of sugar content in the experiment of gas production and lactose decomposition of KM 2017 strain is shown in FIG. In the case of 20% lactose-free samples, the sugar content tended to be retained even after the fermentation period had proceeded at 6.2 ° Brix. On the other hand, in the case of 20% lactose 13 mL and 20% lactose 25 mL, the sample tended to decrease by 5.1 ° Brix and 4.5 ° Brix on the third day of fermentation.

(3) Number of living cells

Kazachstania The results of the viable cell count in the experiment of gas production and lactose decomposition of KM 2017 strain are shown in Fig. The addition of 13 mL of 20% lactose showed a fermentation of 4.41 × 10 ⁵ CFU / mL on the 0th day of fermentation, and the bacterial count tended to rise at 5 × 10 ⁷ CFU / mL on day 1 of fermentation. The number of viable cells in the third day of fermentation tended to be maintained, and the number of bacteria in the fifth day of fermentation decreased to 1.12 × 10 ⁷ CFU / mL. On the other hand, the sample containing 25 mL of 20% lactose showed 7.7 × 10 ⁵ CFU / mL on day 0 of fermentation. Thereafter, the number of bacteria on the first day of fermentation increased and the number of bacteria on the third day of fermentation decreased. The 5th day after fermentation showed 1.6 × 10 ⁷ CFU / mL.

(4) Gas production experiment

Kazachstania The results of gas production in the gas production and lactose decomposition experiments of KM 2017 strain are shown in FIG. In the case of 20% lactose-free samples, no gas was produced. In the case of samples containing 20 mL of lactose 13 mL, it was confirmed that about 0.6 cm of gas was produced 42 hours after fermentation. After 90 hours of fermentation, about 6.8 cm of gas was produced. For the samples added with 20% lactose 25 mL, about 1 cm gas was produced after 36 hours of fermentation. Therefore, it was confirmed that there was a difference in the gas generating ability depending on the amount of lactose added.

< Example  7> Kazakhstania Serbazi ( Kazachstania servazzii ) Confirmation of availability of lactose by the whey concentration of strain KM 2017

1. Experimental Method

As a result of the analysis of general components of whey, 98.24% of moisture, 0.44% of crude fat, 1.16% of crude ash, and protein content of 1.06 ~ 1.28% of whey. The lactose content of whey was about 4 ~ 5%. Experiments were carried out using K. servazzii , which has been confirmed to have lactose resolution, using whey stock solution and two-fold dilution of whey. The whey stock solution and the diluted whey samples were prepared, sterilized at 121 ° C for 15 minutes, inoculated with K. servazzii , and cultured for 5 days (FIG. 30 and Tables 5 to 7).

A B Whey 95 mL 50 mL D.W 0 mL 50 mL 10% yeast extract 5 mL 5 mL Yeast K. servazzii
0.4 mL K. servazzii
0.4 mL

Whey moisture 98.24% Crude fat 0.44% Crude protein 1.16% citric acid 104.80 mg% lactic acid 44.64 mg% glutamic acid 182.73 mg%

Whey pH 6.20 ± 0.01 Acidity (%) 0.18 + 0.03 Soluble solid content (° Brix) 18.2 ± 0.2 Flavonoid (mg / g) 140.21 + - 3.95 Polyphenol (mg / g) 22.43 + - 2.58

2. Experimental results

(1) pH and acidity

Kazachstania servazzii) KM pH and acidity in 2017 of lactose (lactose of the whey concentration) the resolution of the test strains is shown in Figure 31. In the case of the whey crude solution, the fermentation is represented by the pH value of the first day of the test, which is 6.21. The pH tended to be maintained until the first day of fermentation. And thereafter the fermentation tended to decrease slightly to pH 6.04 on the third day of fermentation. And then decreased to pH 5.82 on the 5th day of fermentation. In the case of whey diluted samples, the fermentation showed a pH of 6.09 on the 0th day. And then the pH of the fermentation day 3 was 5.84. And then decreased to 5th day of fermentation pH 5.66. In the case of acidity, fermentation of the two samples tended to slightly increase by 0.20% and 0.23% on the third day of fermentation. The samples of the 5th day after fermentation were 0.25% and 0.29%, respectively.

(2) sugar content

Kazachstania servazzii ) The sugar content in the lactose decomposition test according to the whey concentration of KM 2017 strain is shown in FIG. The whey stock solution tended to decrease slightly starting from 18.2 ° Brix on day 0 of fermentation and 16.2 ° Brix on day 3 of fermentation. And then 13 ° Brix on the 5th day of fermentation. In the case of whey diluted samples, the fermentation showed 3 days of 8.1 ° Brix and 5 days of fermentation of 5.3 ° Brix.

(3) Number of living cells

Kazachstania The number of viable bacteria in the lactose decomposition test according to the whey concentration of KM 2017 strain is shown in FIG. The number of viable cell counts of fermented milk was 2.73 × 10 ⁵ CFU / mL for fermented milk, and the fermentation was increased to 5.03 × 10 ⁷ CFU / mL on day 1 of fermentation. After 10 ⁷ showed a tendency to be kept in CFU / mL are shown the primary fermentation 5 1.52 × 10 ⁷ CFU / mL. For whey-diluted samples, starting with 3.08 × 10 ⁵ CFU / mL. Thereafter, it tended to increase to 5.51 × 10 ⁷ CFU / mL and showed a tendency to maintain the number of viable bacteria at the third day of fermentation. Followed by 3.55 × 10 ⁷ CFU / mL on the fifth day of fermentation.

(4) Gas production experiment

Kazachstania The results of gas production in the experiment of lactose decomposition according to whey concentration of KM 2017 strain are shown in FIG. Each sample did not produce gas even after 36 hours of fermentation. On the other hand, after 48 hours of fermentation, it was confirmed that about 0.2 cm of gas was produced in the diluted sample of whey. After 60 hours of fermentation, it was confirmed that gas was also produced in the whey crude solution. After 90 hours of fermentation, 2.6 cm gas was produced in the stock solution of whey and 4.1 cm gas was produced in the case of diluted whey sample.

That is, as mentioned in the above examples, the ability to degas the lactose of the Kazakhstania servazzii strain KM 2017 was confirmed by performing a gas production ability test and a lactose resolving ability experiment under three different culture conditions. The YPD medium used in the present invention was further tested by addition of lactose to a sugar-free medium. As a result, it was confirmed that the pH was lowered. In addition, it was confirmed that lactose was resolvable by confirming that the sugar content was lowered on the 3rd to 5th days of fermentation. In addition, whey contains about 3 ~ 5% of lactose, and as a result of the resolution test, the sugar content was lowered and the viable cell count was confirmed.

According to previous reports, Kazachstania servazzii strain was found to be unable to utilize lactose. However , the strains of Kazakhstania servazzii ) KM 2017 strain (KCCM11989P) was differentiated from its lactic acid by using sucrose as a fermentable sugar as well as lactose.

Korea Microorganism Conservation Center (overseas) KCCM11989P 20170316

Claims (6)

KM 2017 strain of Kazachstania servazzii isolated from apple pickles, which is entrusted with KCCM 11989P and has excellent lactose resolving power and gas generating ability and has excellent whey efficacy upon whey fermentation. delete delete delete A method of whey fermentation comprising inoculating a strain according to claim 1 into a medium containing whey and incubating at 25 to 30 DEG C for 1 to 5 days for fermentation. delete

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