KR20170040634A - Method for production of maltose with novel enzyme - Google Patents
Method for production of maltose with novel enzyme Download PDFInfo
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Abstract
Lactobacillus plantarum subsp. Plantarum < RTI ID = 0.0 > The present invention relates to a method for producing maltose by using an enzyme having an amino acid sequence of SEQ ID NO: 1 derived from ST-III strain. Since the reaction can be performed even at a low temperature of 30 DEG C, (LBA) is similar to the alpha amarase belonging to the carbohydrate family (GH family 13) or beta amylase which hydrolyzes the substrate from the non-reducing end to the maltose unit, so that it can directly produce maltose have.
Description
The present invention relates to a method for producing maltose using a novel enzyme, and more particularly, to a method for producing maltose using Lactobacillus plantarum subsp. Plantarum ( Lactobacillus plantarum subsp. Plantarum ) A method for producing maltose using an enzyme derived from ST-III strain is provided.
Maltose (malate, G2) is derived from malt, which means malt (malt, malt), also called maltose. Maltose is a reducing disaccharide in which two glucose molecules are linked to an alpha-1,4-bond.
Maltose is one of the maltooligosaccharides with high anabolic activity and high nutritional value. It is usually contained in germinated seeds in the natural world. Such maltose is widely used in industries such as food, pharmacy, biomedicine, and refining chemistry.
For the production of maltose, the method of increase was mainly used. The addition method is a two step process in which a heat-resistant alpha amylase, which is a liquefaction enzyme, is added to starch and reacted at the same time with the growth at 90 to 100 ° C to liquefy, and then various maltose-producing enzymes (beta amylase) are added to the liquefied starch for 24 to 72 hours 'Liquify liquefaction - saccharification process' means.
However, the above-mentioned conventional processes have a disadvantage in that a large amount of energy is consumed and the saccharification time is long in the process of increasing and concentrating. Therefore, it is necessary to develop a new method to solve this problem.
In the present invention, in order to overcome the disadvantage that a large amount of energy is consumed and the saccharification time is long in the conventional step of increasing and concentrating, a new enzyme capable of proceeding the reaction at a low temperature is discovered and a new method for producing maltose is developed I want to.
The present invention provides a method for producing maltose, which comprises treating an enzyme having an amino acid sequence of SEQ ID NO: 1 with a compound having an? -1,4 glucoside bond to react. An example of a compound having an? -1,4 glucoside bond is amylose, and starch may also be present.
Meanwhile, in the present invention, the compound having an? -1,4 glucoside bond may be one having an? -1,6 glucoside bond as an example. The compound having an? -1,6 glucoside bond other than? -1,4 glucoside bond is, for example, amylopectin.
In the present invention, alpha-amylase (LBA) cloned from a Lactobacillus plantarum subsp. Plantarum ST-III strain is used as a starch, and maltose (saccharide), which is a relatively high value- (30 ℃) at a high yield.
Generally, in order to produce maltose from starch, a liquefaction process performed at a high temperature and a saccharification process followed by the saccharification process must be performed together. The LBA enzyme of the present invention can perform both liquefaction and saccharification processes at a low temperature. The fact that the liquefaction reaction is possible at a low temperature means that the energy saving efficiency is higher than the conventional process. From this, the present invention can secure high productivity.
In addition, since the LBA enzyme of the present invention can also carry out a saccharification process for producing maltose from starch hydrolyzate, it is not necessary to use two enzymes. Therefore, the present invention has an advantage that maltose can be produced even by using only one LBA enzyme of the present invention. This can simplify the process and reduce the cost of using the enzyme, thus contributing to an improvement in productivity of maltose production.
On the other hand, the enzyme (LBA) of the present invention can produce maltose with high purity because maltotriose can be degraded unlike the conventional enzymes (made of beta amalia). That is, when a conventional enzyme is used, since maltotriose can not be decomposed, maltotriose exists in addition to maltose in the product. However, since the enzyme (LBA) of the present invention can also decompose Maltotriose to produce maltose, maltose can be produced with high purity.
On the other hand, the present invention relates to a method for producing maltose, which comprises producing maltose from maltotriose by treating an enzyme having the amino acid sequence of SEQ ID NO: 1 with a substrate containing maltotriose, ≪ / RTI >
On the other hand, in the maltose production method of the present invention, the reaction is preferably carried out at a temperature of 10 to 50 ° C. Since the enzyme of the present invention exhibits high activity even at a relatively low temperature, it is not necessary to induce the reaction at a high temperature by consuming a lot of energy.
When the enzyme (amino acid sequence of SEQ ID NO: 1, LBA) derived from Lactobacillus plantarum subsp. Plantarum ST-III of the present invention is used, It has the advantage of saving energy because reaction is possible.
Further, since the enzyme (LBA) of the present invention has properties similar to those of the alpha-amylase belonging to the carbohydrate hydrolase family (GH family, GH) 13 or beta amylase hydrolyzing the substrate from the non-reducing end to the maltose unit, It can directly produce maltose.
Further, since the enzyme (LBA) of the present invention has the specificity to hydrolyze maltotriose (sugar composed of three glucose) which can not be hydrolyzed by general beta amylase, which is an enzyme derived from a plant, There is an advantage that maltose can be produced.
Fig. 1 is a map of a recombinant vector into which insert DNA ( lpst- co1426, the gene of the present invention LBA enzyme) prepared in the present invention is inserted.
2 is an electrophoresis photograph of the purified LBA enzyme of the present invention. 2: S: standard solution, lane 1: cell extract, lane 2: soluble part, lane: purified enzyme of the present invention (LBA).
3 is a TLC photograph showing the substrate degradation pattern of the LBA enzyme of the present invention. 3, maltotriose, lane 3: maltotetraose, lane 4: maltoheptaose, lane 5: alpha -CD, lane 6: beta -CD, lane 7: gamma -CD, lane 8: amylose, lane 9: amylopectin, lane 10: soluble starch; a: before reaction, b: after reaction '.
Fig. 4 is a photograph showing the decomposition pattern of the LBA enzyme of the present invention against p NPG5 (substrate). Fig. Lane 1: p NPG5 control, lane 2:
Fig. 5-A shows the results of experiments in which the optimal reaction temperature of the LBA enzyme of the present invention was confirmed. FIG. 5-B shows experimental results of confirming the optimum pH for the reaction of the LBA enzyme of the present invention.
FIG. 6-A shows the substrate hydrolysis patterns of the LBA enzyme of the present invention and the existing beta-amalase. 6-A: S: G1-G7 standard solution, lane 1: maltotriose, lane 2: maltopentaose, lane 3: maltohexaose; a: before the reaction, b: after the LBA reaction, and c: after the beta amylase reaction. Figures 6-B and C are the results showing the purity of maltose produced by the enzyme LBA of the present invention and conventional beta-amalase treatment. In FIGS. 6-B and C, 'B: LBA reaction, C: beta amylase reaction; a: soluble starch treated with isoamylase, b: 1 hour reaction c: 7 hours reaction d: 12 hours reaction.
Hereinafter, the present invention will be described in more detail with reference to the following Examples and Experimental Examples. However, the scope of the present invention is not limited to the following embodiments and experimental examples, and includes modifications of equivalent technical ideas.
[ Example 1: Lactobacillus Flora Room Subspecies Flora Room ( Lactobacillus plantarum subsp . plantarum ) ST-III LBA Production of enzyme (amino acid sequence of SEQ ID NO: 1)
One) Polymerase chain reaction (Polymerase chain reaction; PCR )
Lactobacillus plantarum subsp. Plantarum < RTI ID = 0.0 > (Having the nucleic acid sequence of SEQ ID NO: 2) of the LBA enzyme of the present invention (having the amino acid sequence of SEQ ID NO: 1) from ST-III strain (KCTC 3015).
Add 37.5 μL of sterilized water, 5 μL of 10 × EX Taq buffer, 4 μL of dNTP, 1 μL of forward primer, 1 μL of reverse primer, DNA (KCTC 3015, Lactobacillus plantarum subsp. Plantarum ST -III) and a 0.5 μL mixture of EX Taq (total 50 μL) were subjected to 1 minute at 98 ° C using the primer shown in Table 1, followed by denaturation at 98 ° C for 10 seconds, PCR (30 cycles) was performed under the conditions of annealing at 72 ° C for 1 min and elongation at 30 ° C.
2) Of the polymerase chain reaction refine
After the PCR reaction, the PCR mixture was vortexed with 5X the volume of BNL buffer. Thereafter, isopropanol, which is 1.5 times the volume of the BNL buffer, was added and pipetted several times to mix. This was transferred to a spin column and centrifuged at 11,323 x g for 1 minute to discard the supernatant. 700 μL of wash buffer was added and centrifuged at 11,323 × g for 1 minute to discard the supernatant. Thereafter, the cells were centrifuged again at 11,323 x g for 1 minute in a state where nothing was added. The top of the spin column was transferred to a new microcentrifuge tube, and 30 μL of sterilized water was added to the microcentrifuge tube. The sample was allowed to stand for 1 minute and then centrifuged at 11323 × g for 1 minute. The concentration of the produced PCR product ( lpst- co146 ) was measured with a nanodrop, which was 215.1 ng / μL.
3) Restrict enzyme treatment
A mixture of 28 μL of the DNA ( lpst- cor146 ) obtained in the above PCR, 2 μL of Sal I, 2 μL of Xho I, 5 μL of 1 × H buffer and 13 μL of sterilized water was reacted at 37 ° C. for 2 hours . As a vector, pTKNd119 was used for the restriction enzyme reaction in the same manner. Then, 1 μL of CIAP was added to the pTKNd119 vector alone, and reacted at 37 ° C. for 1 hour.
4) Gel extraction
5 μL of loading buffer was added to each of the inserts (lpst-c0146) and vector (pTKNd119) which had been treated with restriction enzymes, and 0.5 × TBE buffer, 1.2% agarose gel, Lt; RTI ID = 0.0 > 50 < / RTI > Electrophoretic banding was cut out and weighed in a sterile microcentrifuge tube. BNL buffer as much as three times the weight of the gel was added and dissolved for 10 minutes in a heating block at 55 ° C for 2 to 3 times while vortexing. To this, 1 part by volume of isopropanol was added and pipetted. Each was transferred to a spin column, centrifuged at 11,323 x g for 1 minute, and the lower layer was discarded. 700 μL of washing buffer was added and the mixture was centrifuged at 11323 × g for 1 minute. Then, the lower layer was discarded and centrifuged one more time. Then, the upper part of the spin column was transferred to a microcentrifuge tube, 30 μL of sterilized water was added, and the mixture was centrifuged at 11323 × g for 1 minute. As a result of the concentration measurement, 32.0 ng / μL of the insert and 153.6 ng / μL of the vector were obtained.
5) Ligation (Ligation) and transformation
Ligation (ligation), and self-ligation (control). When ligation is performed, the concentration of the insert and the vector should differ by more than three times. First, 6 μL of insert DNA (192.0 ng / μL), 1 μL of a 4-fold diluted vector (38.4 ng / μL), 0 μL of sterilized water and 7 μL of ligase were added to the ligation mixture. For the control self-ligation mixture, 1 μL of a 4-fold diluted vector, 6 μL of sterile water instead of insert DNA, and 7 μL of ligase were added. The ligation mixture and the self-agitation mixture prepared above were each prepared as a recombinant vector at 16 DEG C for 30 minutes (see Fig. 1). Fig. 1 is a map of a recombinant vector into which insert DNA ( lpst- co1426, the gene of the present invention LBA enzyme) prepared in the present invention is inserted.
Then, using the CaCl 2 method coli; transformant was transformed with the recombinant vector to (E coli MC1061.). For this method, add 200 μL competent cells (MC1061) to each ligation and self-ligation mixture, add 2 μL of ice for 2 min at 42 ° C for 30 min, add 800 μL of LB medium, During the recovery period. Then, 800 μL of the supernatant was discarded by centrifugation at 5,000 × g for 1 minute, and the remaining 200 μL was plated on kanamycin solid medium (LBK) and cultured at 37 ° C. for 12 hours.
6) Cell disruption and purification
Escherichia coli inserted with the lygated vector (experimental group) was inoculated into 1 L of kanamycin liquid medium (LBK), stirred at 150 rpm at 30 캜 for 20 hours in a shaking incubator, and centrifuged at 5,420 × g , 20 minutes). The recovered cells were resuspended in 50 mM citric-NaOH buffer solution (pH 6.0) and then subjected to ultrasonic wave disruption. The crude enzyme solution obtained from the cell disruption was centrifuged (5,420 x g , 20 minutes) to obtain supernatant, which was adsorbed on a Ni-NTA column to obtain the LBA enzyme of the present invention.
SDS-PAGE showed that only the pure LBA was purified (49.9 kDa). Fig. 2 is an electrophoresis photograph of the purified LBA enzyme of the present invention (see Fig. 2). 2, 'S: standard solution, lane 1: cell extract, lane 2: soluble part, lane: purified enzyme of the present invention LBA'.
[ Example 2: enzyme of the present invention ( LBA )]
One) LBA Reaction test by substrate
To investigate the optimum substrate and hydrolytic characteristics of LBA, a 1% substrate (maltose (G2), maltoriose (G3), maltotetraose (G4), maltoheptaose ), Alpha-cyclodextrin (alpha-CD), beta-cyclodextrin (beta-CD), gamma-cyclodextrin (gamma-CD), amylose, Amylopectin, soluble starch) and 0.8 unit / mg LBA enzyme were added to 50 mM NaOAC buffer solution at pH 6.0 and reacted at 30 ° C for 12 hours to perform TLC (thin layer chromatography) analysis.
As a result of TLC, LBA hydrolyzes the substrate to maltose units and does not hydrolyze alpha-CD, beta-CD and gamma-CD (see FIG. 3). 3 is a TLC photograph showing the substrate degradation pattern of the LBA enzyme of the present invention. 3, maltotriose, lane 3: maltotetraose, lane 4: maltoheptaose, lane 5: alpha -CD, lane 6: beta -CD, lane 7: gamma -CD, lane 8: amylose, lane 9: amylopectin, lane 10: soluble starch; a: before reaction, b: after reaction '.
In addition, 0.8 unit / mg of enzyme was added to 50 mM NaOAC buffer solution of 1% p NPG5 and pH 6.0, and reacted at 30 ° C for 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 12 hours And the reaction was stopped by boiling for 5 minutes. As a result of TLC, it was confirmed that G2 and p NPG3 were produced from the early stage of the reaction, and that LBA hydrolyzed the substrate from the non-reducing end (see FIG. 4). Fig. 4 is a photograph showing the decomposition pattern of the LBA enzyme of the present invention against p NPG5 (substrate). Fig. Lane 1: p NPG5 control, lane 2:
2) LBA Optimal temperature and pH confirmation experiment of enzyme
DNS (3,5-dinitrosalicylic acid) was used to determine the optimum temperature and pH of the LBA enzyme of the present invention.
① Check the optimum reaction temperature
The samples were incubated at 10 ~ 50 ℃ for 0.5 min. The samples were then incubated at 5 ~ 10 ℃ for 5 min. The plate was washed with 0.5% soluble starch (showa, Japan), buffer solution (50 mM NaOAC pH6.0) The ratio of the sample to the DNS solution was 1: 3, the mixture was boiled for 5 minutes, and the absorbance was measured at 570 nm using an ELISA reader device. Respectively.
As a result, it was confirmed that the LBA enzyme had the highest activity at 30 ° C (see FIG. 5-A). Fig. 5-A shows the results of experiments in which the optimal reaction temperature of the LBA enzyme of the present invention was confirmed.
② Optimal reaction pH check
(50 mM citrate-NaOH pH 2.5-4, 50 mM NaOAC pH 4-6) and 0.8 unit / mg of LBA enzyme were added to the reaction mixture at a temperature of 30 ° C. After 5 minutes of mixing, the samples were mixed with the DNS solution at a ratio of 1: 3, and the absorbance was measured at 570 nm using an ELISA reader apparatus. And the standard deviation was calculated.
As a result, it was confirmed that the LBA enzyme had the highest activity at pH 3.0 (see FIG. 5-B). FIG. 5-B shows experimental results of confirming the optimum pH for the reaction of the LBA enzyme of the present invention.
3) LBA Enzymatic Wrecker specific activity measurement
Each 0.5% substrate (maltotriose, maltotetraose, maltopentaose, glycogen, potato starch, corn starch, soluble starch, , Amylose and amylopectin) and 0.8 unit / mg of LBA enzyme were added to 50 mM citric-NaOH buffer solution of pH 3.0, and reacted at 30 ° C. for 3 minutes, 6 minutes, 9 minutes, 12 minutes and 15 minutes Min and the ratio of sample to DNS was 1: 3. After boiling for 5 minutes, the absorbance was measured at 570 nm using an ELISA reader device. The standard deviations were obtained and the results are shown in Table 2.
As a result, it was confirmed that the LBA enzyme exhibited the best non-reactivity of 3.15 ± 0.110 U / mg in maltopentaose.
4) LBA Reactivity of Enzymes ( k cat / K m ) Kinetic study
Add 50 mM Citric-NaOH pH 3.0 and 0.8 unit / mg of LBA enzyme to each substrate (glycogen, potato starch, corn starch, amylose, amylopectin) ℃ and samples are sampled every 3, 6, 9, 12, 15, and 18 minutes and boiled for 5 minutes. Each substrate was tested at various concentrations.
Analysis of the samples was performed using high performance anion exchange chromatography (HPAEC). The LBA enzyme reaction solution was diluted 50 times with distilled water, filtered through a membrane filtration filter (0.45 mu m), and then 20 μL was injected into the instrument and the flow rate was 1 mL / min. HPAEC analysis was carried out using Dionex's GP40 gradient pump and ED40 electrochemical detector and Carbo-PacPA1 column. A 150 mM NaOH solution was used as the solvent A, 600 mM sodium acetate (Na-acetate) was dissolved in the solvent A, and B solvent was changed from 10% to 22% after 6 minutes. The concentration was increased linearly. Table 3 shows the results.
The results were as follows: k cat / K m (mL / mg · min), glycogen 0.888 mL / mg · min, potato starch 0.912 mL / mg · min, corn starch 0.498 mL / mg · min, amylose 1.164 mL / mg · min, and amylopectin: 0.714 mL / mg · min.
5) LBA and Comparison of existing plant-derived β-amylase (plant origin β-amylase)
For comparison of the new enzyme LBA with the conventional beta amylase (commercially available enzyme-beta amylase from barley, Sigma-Aldrich Co.), 0.5% substrate (maltotriose, maltopentaose, maltohexaose) , 50 mM citric-NaOH buffer solution and 0.8 unit / mg of enzyme were mixed and reacted at 30 ° C for 12 hours. In the case of beta amylase, a 50 mM NaOAc buffer solution of pH 5.0 and 0.8 unit / mg of enzyme were mixed And reacted at 20 ° C for 12 hours. Thereafter, the reaction was stopped by boiling for 5 minutes, followed by TLC (see FIG. 6-A). Fig. 6-A shows the substrate hydrolysis patterns of the enzyme LBA of the present invention and the existing beta-amalase. 6-A: S: G1-G7 standard solution, lane 1: maltotriose, lane 2: maltopentaose, lane 3: maltohexaose; a: before the reaction, b: after the LBA reaction, and c: after the beta amylase reaction.
As a result, it was confirmed that the existing β-amylase hydrolyzes G6 and G5 in maltose unit, but G3 does not hydrolyze. However, in the case of LBA, not only G6 and G5 were hydrolyzed in maltose units, but also G3 was hydrolyzed.
In order to compare the purity of maltose produced with each enzyme, 0.5% soluble starch was treated with 50 mM NaOAC pH 4.0, 0.2 unit / mg of isoamylase for 4 hours at 40 ° C, The amount of enzyme was 0.5 unit / mg. LBA was reacted at 50 ℃ in Citric-NaOH pH 3.0 at 30 ℃, and 50 mM NaOAC pH 5.0, 20 ℃ in beta amylase. After 1 hour, 7 hours, and 12 hours, they were sampled and boiled for 5 minutes to stop the reaction.
Then, the LBA enzyme and beta amylase reaction solution was diluted 50 times with distilled water and filtered with a thin membrane filtration filter (0.45 mu m), and then 20 μL was injected into the device and the flow rate was 1 mL per minute. HPAEC analysis was carried out using Dionex's GP40 gradient pump and ED40 electrochemical detector and Carbo-PacPA1 column. A 150 mM NaOH solution was used as the solvent A, and 600 mM sodium acetate (Na-acetate) was dissolved in the solvent A, which was used as the solvent B. After 16 minutes, the solvent B was changed from 10% to 40% And the concentration was linearly increased (see Figs. 6-B and C). Figures 6-B and C are the results showing the purity of maltose produced by the enzyme LBA of the present invention and conventional beta-amalase treatment. In FIGS. 6-B and C, 'B: LBA reaction, C: beta amylase reaction; a: soluble starch treated with isoamylase, b: 1 hour reaction c: 7 hours reaction d: 12 hours reaction.
As a result of the experiment, LBA hydrolyzes G3, so only the G2 graph can be confirmed. However, the existing beta amylase did not hydrolyze G3, and the graph of G3 was confirmed.
As a result, it was confirmed that, when LBA of the present invention was used, G2 production was possible with high purity as compared with the existing beta amylase.
<110> Industry Academic Cooperation Foundation, Hallym University <120> Method for production of maltose with novel enzyme <130> AP-2015-0165 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 440 <212> PRT <213> Lactobacillus plantarum subsp. plantarum ST-III (KCTC 3015) <400> 1 Met Ala Arg Asp Thr Gln Thr Gln Leu Arg Asn Glu Met Ile Tyr Ser 1 5 10 15 Val Phe Val Arg Asn Tyr Ser Glu Ala Gly Asn Phe Ala Gly Val Thr 20 25 30 Ala Asp Leu Gln Arg Ile Lys Asp Leu Gly Thr Asp Ile Leu Trp Leu 35 40 45 Leu Pro Ile Asn Pro Ile Gly Glu Val Asn Arg Lys Gly Thr Leu Gly 50 55 60 Ser Pro Tyr Ala Ile Lys Asp Tyr Arg Gly Ile Asn Pro Glu Tyr Gly 65 70 75 80 Thr Leu Ala Asp Phe Lys Ala Leu Thr Asp Arg Ala His Glu Leu Gly 85 90 95 Met Lys Val Met Leu Asp Ile Val Tyr Asn His Thr Ser Pro Asp Ser 100 105 110 Val Leu Ala Thr Glu His Pro Glu Trp Phe Tyr His Asp Ala Asp Gly 115 120 125 Gln Leu Thr Asn Lys Val Gly Asp Trp Ser Asp Val Lys Asp Leu Asp 130 135 140 Tyr Gly His His Glu Leu Trp Gln Tyr Gln Ile Asp Thr Leu Leu Tyr 145 150 155 160 Trp Ser Gln Phe Val Asp Gly Tyr Arg Cys Asp Val Ala Pro Leu Val 165 170 175 Pro Leu Asp Phe Trp Leu Glu Ala Arg Lys Gln Val Asn Ala Lys Tyr 180 185 190 Pro Glu Thr Leu Trp Leu Ala Glu Ser Ala Gly Ser Gly Phe Ile Glu 195 200 205 Glu Leu Arg Ser Glu Gly Tyr Thr Gly Leu Ser Asp Ser Glu Leu Tyr 210 215 220 Gln Ala Phe Asp Met Thr Tyr Asp Tyr Asp Val Phe Gly Asp Phe Lys 225 230 235 240 Asp Tyr Trp Gln Gly Arg Ser Thr Val Glu Arg Tyr Val Asp Leu Leu 245 250 255 Gln Arg Gln Asp Ala Thr Phe Pro Gly Asn Tyr Val Lys Met Arg Phe 260 265 270 Leu Glu Asn His Asp Asn Ala Arg Met Met Ser Leu Met His Ser Lys 275 280 285 Ala Glu Ala Val Asn Asn Leu Thr Trp Ile Phe Met Gln Arg Gly Ile 290 295 300 Pro Leu Ile Tyr Asn Gly Gln Glu Phe Leu Ala Glu His Gln Pro Ser 305 310 315 320 Leu Phe Asp Arg Asp Thr Met Val Ala Asp Arg His Gly Asp Val Thr 325 330 335 Pro Leu Ile Gln Lys Leu Val Thr Ile Lys Gln Leu Pro Leu Leu Arg 340 345 350 Ala Ala Asp Tyr Gln Leu Ala Val Val Glu Glu Gly Ile Val Lys Ile 355 360 365 Thr Tyr Arg Ala Gly Glu Ala Leu Thr Ala Trp Ile Pro Leu Lys 370 375 380 Gly Gln Val Thr Ala Val Ala Thr Lys Leu Ala Ala Gly Ser Tyr Gln 385 390 395 400 Asn Leu Leu Thr Asp Gly Pro Thr Glu Val Asp Gly Lys Leu Thr 405 410 415 Val Asp Gly Gln Pro Val Leu Ile Lys Tyr Val Thr Asn Thr Ala Val 420 425 430 Thr Lys Val Ala Asp Gln Ser Asn 435 440 <210> 2 <211> 1323 <212> DNA <213> Lactobacillus plantarum subsp. plantarum ST-III (KCTC 3015) <400> 2 atggcacgcg atacgcaaac gcaattacgc aatgagatga tttactcagt ctttgttaga 60 aactctcag aagctggtaa ttttgctggc gtaactgctg acttacaacg aattaaggat 120 ttaggaaccg atattttgtg gctactgcca atcaatccga tcggtgaggt caaccgtaag 180 gggacacttg gttcgccata tgcaatcaag gactaccgtg ggatcaatcc ggaatacggg 240 actttagctg actttaaagc actcacggat agggcgcatg aattaggaat gaaagtgatg 300 ttagacattg tctataatca cacttcgcct gattcagtct tagcaaccga acatccagag 360 tggttttatc acgacgcgga tggtcagttg acgaataagg tcggcgactg gagtgacgtt 420 aaagacttag actatggtca ccatgagttg tggcagtatc aaatcgacac actcttgtat 480 tggagccagt ttgtggacgg ttaccgttgt gatgttgcgc cattagtgcc acttgatttc 540 tggcttgaag cgcggaaaca ggttaatgcg aagtacccgg aaacgttatg gttagcggag 600 tcagccggca gtggttttat tgaggaactg cggtcacaag gctacacggg tctgtctgac 660 agtgaacttt atcaagcttt tgatatgaca tacgattacg acgtcttcgg tgattttaaa 720 gattactggc aaggacgcag tacggtcgaa cggtatgttg acttgttaca acgccaagat 780 gccactttcc ctggtaatta tgtcaagatg cgcttcttgg aaaatcatga taatgcacgc 840 atgatgagct tgatgcacag caaagctgaa gccgttaata acttgacctg gatcttcatg 900 caacgtggta ttccgttaat ctataacggg caagaatttt tggccgaaca ccaaccatca 960 ttgttcgatc gggacaccat ggttgcagat cgtcatgggg acgtgacacc actgattcaa 1020 aagctagtga ctattaagca actaccatta ctgcgtgctg cggactacca attggcagtc 1080 gtagaagaag gtatcgttaa gattacttac cgtgcggctg gcgaagcgtt aacagcctgg 1140 attccgttaa aaggtcaggt cacagcagtt gcgactaagc tagcagctgg tagctatcaa 1200 aatctgttaa cagatggtcc tactgaagtt gtggatggca agctgacagt tgatggtcaa 1260 ccagtattaa ttaaatatgt caccaatacg gcggtgacta aagttgctga ccagtccaat 1320 tag 1323
Claims (7)
The compound having an? -1,4 glucoside bond,
wherein the maltose has an? -1,6 glucoside bond.
The compound having an? -1,4 glucoside bond,
Wherein the maltose is amylose.
The compound having an? -1,6 glucoside bond in addition to the? -1,4 glucoside bond,
Wherein the amylopectin is amylopectin.
The compound,
Production method of maltose characterized by being starch
The above-
Lt; RTI ID = 0.0 > 10-50 C. < / RTI >
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20200029163A (en) * | 2018-09-10 | 2020-03-18 | 한림대학교 산학협력단 | Method for optimum production of LpMA with Lactobacillus plantarum |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR980002268A (en) | 1996-06-28 | 1998-03-30 | 박관화 | A maltogenic amylase gene, a recombinant molecule containing it, a transformant having the same, and an enzyme produced therefrom |
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| KR980002268A (en) | 1996-06-28 | 1998-03-30 | 박관화 | A maltogenic amylase gene, a recombinant molecule containing it, a transformant having the same, and an enzyme produced therefrom |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200029163A (en) * | 2018-09-10 | 2020-03-18 | 한림대학교 산학협력단 | Method for optimum production of LpMA with Lactobacillus plantarum |
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