KR100460605B1 - Production of the flavor-enhancer by fermentation - Google Patents
Production of the flavor-enhancer by fermentation Download PDFInfo
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- KR100460605B1 KR100460605B1 KR10-2001-0041775A KR20010041775A KR100460605B1 KR 100460605 B1 KR100460605 B1 KR 100460605B1 KR 20010041775 A KR20010041775 A KR 20010041775A KR 100460605 B1 KR100460605 B1 KR 100460605B1
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/20—Synthetic spices, flavouring agents or condiments
- A23L27/24—Synthetic spices, flavouring agents or condiments prepared by fermentation
Abstract
4-에틸 구아이아콜(4-ethyl guaiacol)은 간장, 과실주 등의 발효식품에 미량(3ppm이하) 함유되는 향미성분으로서, 그 향미는 양호하고, 역치는 낮다. 그러므로 4-에틸 구아이아콜의 함량이 높은 향미액(향미료)을 발효법에 의하여, 간이하게 제조할 수 있게 되면, 그 향미액은 식품제조 또는 가공에 있어서, 다양하게 사용될 수 있다. 그러나 4-에틸 구아이아콜의 함량이 높은 향미액의 제조나 그 이용에 관하여는 전혀 알려진 것이 없다. 본 발명은 위와 같은 관점에 따라, 시도하게 된 것이다.4-ethyl guaiacol is a flavor component (3 ppm or less) contained in fermented foods such as soy sauce and fruit wine, and the flavor is good and the threshold is low. Therefore, when a flavor liquid (flavor) having a high content of 4-ethyl guai alcohol can be easily produced by the fermentation method, the flavor liquid can be used in various ways in food production or processing. However, nothing is known about the preparation or use of flavored liquids with a high content of 4-ethyl guiacol. The present invention has been made in accordance with the above aspects.
본 발명은 4-에틸 구아이아콜의 함량이 높은 향미액의 제조에 관한 것이며, 상세하게는 화본과곡류의 곡피 또는 겨에서, 페룰산(ferulic acid)유리효소의 작용으로, 페룰산을 유리(遊離)시켜, 페룰산액을 조제하고, 이 페룰산액에, 페룰산을 4-에틸 구아이아콜로 변환시키는 능력이 높은 칸디다속 효모(Candida versatilis)의 배양물을 첨가하고, 교반 또는 통기교반하면서, 발효시키는 것에 의하여, 4-에틸 구아이아콜의 함량이 높은 향미액을 제조하는 것이다.The present invention relates to the preparation of a flavor liquid having a high content of 4-ethyl guai alcohol, and specifically, in the grains or bran of the flower and grains, ferulic acid (ferulic acid) free enzymes, the release of ferulic acid (遊離) The ferulic acid solution was prepared, and the ferulic acid solution was fermented by adding a culture of Candida versatilis having a high ability to convert ferulic acid to 4-ethyl guacol, and stirring or aeration stirring. By making it, the flavor liquid with high content of 4-ethyl guiacol is produced.
이를 위하여, 본 발명에서는 발효시험에 의하여 선발한, 4-에틸 구아이아콜 생성능이 높은 균주를 발효에 사용하고, 또 발효 중에는 교반 또는 통기교반하는 것에 의하여, 4-에틸 구아이아콜의 생성반응을 효율적으로 진행시키는 것을 특징으로 하였다.To this end, in the present invention, 4-ethyl guai alcohol, which is selected by the fermentation test, is used for fermentation, and the production reaction of 4-ethyl guai alcohol is carried out by stirring or aeration with aeration during fermentation. It was characterized by progressing efficiently.
Description
본 발명은 4-에틸 구아이아콜(4-ethyl guaiacol)의 함량이 높은 향미액 즉, 향미료를 발효에 의하여 제조하는 방법에 관한 것이며, 상세하게는, 화본과곡류의 곡피나 겨에서, 페룰산(ferulic acid)을 유리(遊離)시켜 페룰산액을 조제하고, 농도가 높은 페룰산액에서도, 페룰산을 4-에틸 구아이아콜로 변환시키는 능력이 높은 칸디다속 효모(Candida versatilis) 균주를 사용하여, 4-에틸 구아이아콜을 높은 수율로 생성시키는 방법에 관한 것이다.The present invention relates to a method of preparing a flavor liquid having a high content of 4-ethyl guaiacol, that is, a spice by fermentation, and specifically, in a grain or bran of a flower and cereal grains, ferulic acid ( Ferulic acid solution was prepared by freeing ferulic acid, and Candida versatilis strains having high ability to convert ferulic acid to 4-ethyl guai alcohol, even in high concentration ferulic acid solution, were used. It relates to a process for producing ethyl guiacol in high yield.
4-에틸 구아이아콜은 과실주(사과주, 포도주 등)나 간장류의 제조시, 장기간의 발효·숙성 중에 미량(0∼4ppm) 생성되며 식품의 향미형성에 기여하는 향미성분으로 알려져 있다. 특히 간장류에서, 4-에틸 구아이아콜의 존재는 품질의 우위성을 나타내는 특징적인 향미성분으로 인정되고 있다. 그러므로 간장류 제조업체 및 관련기관에서는, 간장 또는 간장유사 조미액의 제조에 있어서, 4-에틸 구아이아콜을 효과적으로 생성시켜, 품질을 향상시키는 연구가 계속 이루어져 왔다.4-ethyl guiacol is known to be a flavor component that contributes to the flavor formation of foods, produced in trace amounts (0 to 4 ppm) during long-term fermentation and ripening during the production of fruit wine (apple wine, wine, etc.) or soy sauce. In soy sauces, in particular, the presence of 4-ethyl guai alcohol has been recognized as a characteristic flavor component showing superiority in quality. Therefore, studies have been made at the soy sauce manufacturers and related organizations to effectively produce 4-ethyl guai alcohol and to improve quality in the preparation of soy sauce or soy sauce-like seasoning.
종래, 간장제조에 있어서, 4-에틸 구아이아콜 생성능을 갖는 칸디다속 효모의 배양물을, 간장덧 과정에서, 초기에 첨가하여 발효시키면, 4-에틸 구아이아콜의 생성이 뚜렷하고, 4-에틸 구아이아콜을 3ppm 정도 생성시킬 때, 간장의 향미가 양호하게 된다는 연구(일본 장유연구소 잡지, Vol. 8, p.108∼116, 1982년), 간장제조에 있어서, 간장국(麴)을 담금할 때, 담금용수 또는 담금염수 대신에, 내염성 칸디다속 효모의 저염(低鹽)배양액과 에탄올 생성능을 갖는 효모를 첨가하여 향미가 양호한 저염 간장유사 조미액을 제조하는 방법(일본 공개특허공보, 평 4-311368)등이 발표되었다.Conventionally, in the production of soy sauce, when a culture of candida yeast having 4-ethyl guai alcohol producing ability is added and fermented at an early stage during soy sauce addition, 4-ethyl guai alcohol is clearly produced and 4-ethyl guai alcohol is produced. Soy sauce flavor is soaked when producing about 3 ppm of guai alcohol (Japanese Soymilk Research Institute Magazine, Vol. 8, p. 108 to 116, 1982). Instead of immersion water or immersion saline, low salt culture medium of saline resistant Candida yeast and yeast having ethanol-producing ability are added to produce a low-salt soy sauce-like seasoning with good flavor (Japanese Patent Laid-Open No. 4, H4) -311368).
이 밖에 간장 유사조미액을 재조하기 위하여, 간장원료를 국균효소로 가수분해한 분해액(발효원액)을, 간장발효미생물을 균종별로, 고정화한 고정화균체에 차례로 접촉시켜, 조미액을 제조하는 기술도 연구·개발되었는데, 그 내용에는 내염성 칸디다속 효모의 고정화균체에 의하여, 4-에틸 구아이아콜을 생성시키는 공정에 관한 연구도 포함되어 있다.In addition, in order to prepare a soy sauce-like seasoning solution, a technique for preparing a seasoning solution is obtained by contacting a decomposed solution (fermented stock solution) obtained by hydrolyzing soy sauce raw materials with a bacterium enzyme, and then contacting fermented soybean microorganisms with immobilized immobilized cells. It has been developed, including the study of the process for producing 4-ethyl guai alcohol by immobilized cells of flame-resistant candida yeast.
상기한 간장류의 제조 또는 다른 발효식품의 제조에 있어서, 제조과정 중에 생성되는 4-에틸 구아이아콜의 양은 적고, 간장류의 경우도 그 양은 3ppm이하가 보통이다. 그러나 4-에틸 구아이아콜의 함량이 높은 향미액이나 유사한 제품에 대하여는 전혀 알려져 있지 않다. 그러므로 4-에틸 구아이아콜의 함량이 높은 향미액(향미료)을 효율적으로 제조하는 방법과 그 이용에 관한 연구·개발이 요망된다.In the production of the above soy sauces or other fermented foods, the amount of 4-ethyl guiacol produced during the manufacturing process is small, and in the case of soy sauce, the amount is usually 3 ppm or less. However, no known flavors or similar products with high content of 4-ethyl guaric are known. Therefore, it is desirable to research and develop a method for efficiently preparing a flavor liquid (flavour) having a high content of 4-ethyl guiacol and its use.
본 발명은 저렴한 페룰산 원료에서, 고농도의 페룰산액을 쉽게 제조하는 방법, 그리고 조제한 페룰산액에 4-에틸 구아이아콜 생성능이 높은 칸디다속 효모의 배양물을 첨가하여 발효시키는 것에 의하여, 4-에틸 구아이아콜을 효율적으로 생성시키는 방법을 제공하는데 목적이 있다.The present invention relates to a method of easily preparing a high concentration of ferulic acid liquid from an inexpensive ferulic acid raw material, and to fermenting by adding a fermented yeast culture with high 4-ethyl guai alcohol production ability to the prepared ferulic acid liquid. An object of the present invention is to provide a method for efficiently generating guiacol.
본 발명의 수행에 있어서는, 우선 페룰산함량이 높은 발효원료액에서, 4-에틸 구아이아콜을 효율적으로 생성할 수 있는 유용균주를 얻어야 하는데, 칸디다속 효모의 4-에틸 구아이아콜 생성능은 균주에 따라 차이가 있고, 또 발효원료액중의 고농도의 페룰산은 4-에틸 구아이아콜 생성균에 대하여 영향을 미치므로, 4-에틸 구아이아콜 생성균을 사용하드라도, 4-에틸 구아이아콜의 생성능이 상당히 높은 균주가 아니면, 발효시에 4-에틸 구아이아콜의 생성반응이 지연되거나, 반응이 충분히 진행되지 못하여, 소기의 목적을 이룰 수가 없었다. 따라서 본 발명의 방법에서는, 4-에틸 구아이아콜 생성능이 특히 높은 균주를 검색하여 사용하고, 발효원료액의 페룰산농도를 알맞게 조절하여 사용하는 것 등을 기술적과제로 하였다.In the practice of the present invention, first, a useful strain capable of efficiently producing 4-ethyl guai alcohol in fermented feedstock with high ferulic acid content should be obtained. In addition, the high concentration of ferulic acid in the fermentation feedstock affects 4-ethyl guai alcohol producing bacteria, so even if 4-ethyl guai alcohol producing bacteria is used, the production ability of 4-ethyl guai alcohol is reduced. If the strain was not very high, the production reaction of 4-ethyl guiacol during fermentation was delayed, or the reaction did not proceed sufficiently, and thus could not achieve its intended purpose. Therefore, in the method of the present invention, a strain having a particularly high 4-ethyl guai alcohol producing ability was searched for and used to adjust the ferulic acid concentration of fermented feedstock accordingly.
본 발명은 화본과곡류의 곡피 또는 겨를 원료로 하여, 4-에틸 구아이아콜(4-ethyl guaiacol)의 함량이 높은 향미액(향미료)을 제조하는 것이며, 상세하게는, 곡류의 곡피 또는 겨에서 국균(Aspergillus oryzae)의 효소작용에 의하여, 페룰산(ferulic acid)을 유리(遊離)시켜 페룰산액을 조제하고, 이 페룰산액에 4-에틸 구아이아콜 생성능이 높은 칸디다속 효모(Candida versatilis)의 배양액을 첨가하여 발효시킴으로서, 4-에틸 구아이아콜의 함량이 높은 발효액 즉, 향미액을 얻는 것이다. 본 발명의 방법을 공정별로 설명하면 다음과 같다.The present invention is to prepare a flavourant (flavour) having a high content of 4-ethyl guaiacol (grain) from grains and bran of flower and grains, and in detail, Ferulic acid is prepared by enzymatic action of Aspergillus oryzae to prepare a ferulic acid solution, and a culture solution of Candida versatilis having high ability to produce 4-ethyl guai alcohol is prepared in this ferulic acid solution. By fermentation by the addition, fermentation broth with a high content of 4-ethyl guai alcohol, that is, flavoring liquid is obtained. The process of the present invention will be described below step by step.
(1) 페룰산액의 조제(1) Preparation of Ferulic Acid Liquid
페룰산액의 조제에 있어서는, 페룰산을 결합형으로 함유하고 있는 밀기울, 쌀겨, 옥수수의 곡피 등을 유용한 원료로 사용할 수 있음으로, 본 발명에서는 페룰산 원료인 밀기울에 국균(Aspergillus oryzae)를 배양한 국균효소제를 작용시키는 방법을 사용하였는데, 이때에는, 효소제에 함유되어 있는 페룰산유리효소의 작용에 의하여 페룰산이 유리될 뿐만 아니라, 이 효소작용을 강화하는 다른 효소(xylanase, arabinosidae 등)도 작용하게 되어 페룰산의 유리·생성량이 크게 증가하게 된다.In the preparation of ferulic acid liquid, bran, rice bran, corn grains, and the like containing ferulic acid in a bound form can be used as a useful raw material, and in the present invention, aspergillus (Aspergillus oryzae) is cultured on bran, which is a ferulic acid raw material. This method was used to act as a bacterium enzyme. At this time, not only ferulic acid is released by the action of the ferulic acid free enzyme contained in the enzyme, but also other enzymes (xylanase, arabinosidae, etc.) that enhance this enzyme action. The amount of glass and production of ferulic acid is greatly increased.
그러므로 페룰산액을 조제할 때에는, 실제의 사용량 시험에 의하여 페룰산 원료인 밀기울과 국균효소제의 적당한 사용량을 사용량비율로서 정하였다. 페룰산 원료인 증자한 밀기울과 국균효소제를 약 9:1 (페룰산 원료는 증자전의 밀기울 무게로, 국균효소제는 건조한 것의 무게로 환산함)로 사용하였으며, 이들의 사용량을 합친 것을, 이것의 3∼5배량 (W/W)의 약 50℃의 온수에 혼합하고, 45℃로 3∼6시간, 페룰산의 추출처리를 한 뒤, 간단한 여과로 고형물을 제거하여, 페룰산액을 얻었다.Therefore, when preparing the ferulic acid liquid, an appropriate amount of fermented wheat bran and a bacterium enzyme was used as the usage ratio by the actual amount test. The increased amount of fermented wheat bran and the bacterium enzyme was used for about 9: 1 (the ferulic acid ingredient was converted to the weight of bran before cooking and the bacterium enzyme was converted into the weight of the dried one). The mixture was mixed with hot water at approximately 50 ° C. of ˜5 times (W / W), extracted with ferulic acid at 45 ° C. for 3 to 6 hours, and then the solid was removed by simple filtration to obtain a ferulic acid solution.
페룰산액의 조제에 있어서, 1차 추출만으로 얻는 페룰산액의 페룰산 함량이 부족할 때에는, 1차 추출로 얻는 페룰산액에 다시, 새로운 페룰산 생성원료(증자한 페룰산 원료와 국균류효소제)를 가하여, 페룰산추출을 되풀이하는 방법(2차 또는 3차 추출하는 방법)으로, 페룰산함량이 높은 페룰산액을 조제한다.상기와 같이 조제한 페룰산액은 pH를 4.6∼5.0으로 조정하고, 가열처리한 뒤에 사용한다.In the preparation of ferulic acid liquid, when the ferulic acid content of the ferulic acid liquid obtained only by primary extraction is insufficient, the ferulic acid liquid obtained by the primary extraction is added again with a new ferulic acid-producing raw material (produced ferulic acid raw material and a national bacterium enzyme). The ferulic acid solution having a high ferulic acid content is prepared by a method of repeatedly extracting ferulic acid (second or third extraction). The ferulic acid solution prepared as described above is adjusted to pH 4.6 to 5.0 and heated. use.
(2) 칸디다속 효모의 배양(2) Cultivation of Candida Yeast
칸디다속 효모의 배양에 있어서는, 발효시험에 의하여 선발한, 4-에틸 구아이아콜 생성능이 높은 균주(NRRL Y-6652균)를 사용하고, 상법으로 제조한 각종 국균의 국(麴) 당화액 또는 엿기름 당화액등을 배지로 사용하였는데, 당화액 배지는 pH를 4.6∼5.0으로 조정하고 가열살균한 후에 사용하였다.In cultivation of candida yeasts, the saccharification liquid of various Korean bacteria produced by the conventional method using a strain having high 4-ethyl guai alcohol production ability (NRRL Y-6652) selected by the fermentation test, or Malt saccharification liquid was used as a medium, and saccharification liquid medium was used after adjusting the pH to 4.6 to 5.0 and heat sterilization.
흑국균의 변이주(Aspergillus awamorivar.kawachii)를 배양한 국의 당화액(산성배지임)은 pH를 조정하지 않고 그대로 사용하였다. 이들 배지외에 선정효모의 배양에 있어서는 필요에 따라 공지의 효모균배지를 사용할 수도 있다.칸디다속 효모를 배양할 때에는, 상법에 따라 배양온도 26∼30℃로 종배양하고, 적당히 확대배양하여 사용하는 데, 배양방법은 진탕배양, 교반배양, 통기교반배양 중에서 적의 선택한다.The saccharification solution (acid medium) of the culture in which the strain of Aspergillus awamori var.kawachii was cultured was used as it was without adjusting the pH. In addition to these mediums, known yeast cell cultures may be used, if necessary, for the cultivation of selected yeasts. When cultivating candida yeasts, they are cultivated at a culture temperature of 26 to 30 ° C. according to a conventional method, and used for expansion and expansion. , The culture method is selected from the shaking culture, stirred culture, aeration stirring culture.
(3) 4-에틸 구아이아콜 발효(3) 4-ethyl guai alcohol fermentation
4-에틸 구아이아콜 발효는 페룰산이 탈탄산(脫炭酸)되고, 이어서 환원(수소부가)되는 2단계의 효소반응에 의하여 이루어지는데, 이 효소반응은 4-에틸 구아이아콜 생성능이 높은 칸디다속 효모에 의하여 효율적으로 진행될 수 있다. 따라서 조제한 페룰산액에 선정한 효모균주의 배양물을 첨가한 뒤, 이것을 교반 또는 통기 교반하면서, 28∼30℃로 1.5∼2.5일간 발효시킨다. 발효 과정 중의 교반 또는 통기교반은 칸디다속 효모의 4-에틸 구아이아콜 생성 반응을 촉진하고, 또 발효액을 칸디다속 효모의 생육에 알맞는 호기상태를 조성하기 위한 것이다. 그러므로 발효탱크의 용량이 큰 경우에는 통기교반에 의한 발효법이 필요하게 된다.Fermentation of 4-ethyl guai alcohol is carried out by a two-step enzymatic reaction in which ferulic acid is decarbonated and then reduced (hydrogenated). This enzyme reaction is a candida genus having high ability to produce 4-ethyl guai alcohol. By yeast can proceed efficiently. Therefore, after adding the culture of the yeast strain selected to the prepared ferulic acid solution, it is fermented at 28 to 30 ° C. for 1.5 to 2.5 days with stirring or aeration. Stirring or aeration agitation during the fermentation process promotes the 4-ethyl guai alcohol production reaction of the Candida yeast, and the fermentation broth to create an aerobic state suitable for the growth of Candida yeast. Therefore, when the capacity of the fermentation tank is large, the fermentation method by aeration stirring is required.
발효가 종료된 후, 발효액은 4-에틸 구아이아콜 함량이 높고, 향미가 양호한 향미액이 되는데, 이 향미액은 여과하고 가열처리하여, 그대로 또는 가공처리한 후 사용한다.After the fermentation is completed, the fermentation broth is a flavor liquid having a high content of 4-ethyl guai alcohol and having a good flavor. The flavor solution is filtered and heated, and is used as it is or after being processed.
(실시 예 1) 향미액의 제조Example 1 Preparation of Flavor Liquid
(1) 페룰산액의 조제(1) Preparation of Ferulic Acid Liquid
밀기울에 100%량 (W/W)의 물을 혼합하고, 120℃로 60분간 가압증자한 것을 페룰산원료로 사용하였으며, 페룰산유리효소로서는, 위와 같이 가수하여 증자한 밀기울에 국균(Aspergillus oryzae)의 종균을 접종하고, 30℃로 2일간 배양한 국균효소제(국균을 배양한 국)를 사용하였다.페룰산액의 조제에 있어서는, 증자한 밀기울(증자전의 밀기울의 무게로서)과 국균효소제(건조물의 무게로서)를 9:1 정도로 사용하는 것이 시험결과 적당하였다.100% (W / W) of water was mixed with bran and pressurized steam at 120 ° C. for 60 minutes was used as ferulic acid raw material. As ferulic acid free enzyme, aspergillus oryzae was added to the bran hydrolyzed as above. Bacillus seed was inoculated and the Bacillus enzyme (Korean culture incubated) was incubated at 30 ° C. for 2 days. In preparation of ferulic acid solution, increased wheat bran (as weight of wheat bran before steaming) and Bacillus enzyme (dry matter) A test weight of 9: 1) was adequate.
그러므로, 2ℓ삼각플라스크에 50℃의 온수 800㎖을 넣고, 이것에 상기한 증자밀기울 180g과 국균효소제 20g을 가하여, 45℃로 3시간 페룰산을 추출한 뒤, 간단한 여과로 고형물을 제거하여, 페룰산액을 조제하였는데, 1차 추출한 페룰산액에 다시 위와 동일하게, 증자한 밀기울과 국균효소제를 가하여, 페룰산을 추출하는 방법을 되풀이하여, 2차 추출 또는 3차 추출한 페룰산액을 조제하였다.페룰산액의 수량과 페룰산농도를 표 1에 표시하였다. 페룰산액의 pH는 어느 것이나 6.0∼6.2 였으며, 페룰산액은 모두 식품용 구연산을 첨가하여, pH를 4.6으로 조정한 후에 사용하였다.페룰산액 중에는 페룰산 외에 적은양의 4-비닐 구아이아콜이 함유되는데, 4-비닐 구아이아콜은 페룰산에서 4-에틸 구아이아콜이 생성되는 과정의 중간산물이며, 이것은 페룰산과 함께 4-에틸 구아이아콜의 원료가 된다. 그러므로 본 발명에서는, 4- 에틸 구아이아콜의 실제원료를 페룰산량으로 표시하기 위하여, 페룰산량에 4- 비닐 구아이아콜량을 페룰산으로 환산한 양을 합하여, 페룰산량으로 표시하였다.Therefore, 800 ml of hot water at 50 ° C. was added to a 2 l triangle flask, and 180 g of the above-mentioned wheat bran and 20 g of the bacterium enzyme were added thereto. The ferulic acid was extracted at 45 ° C. for 3 hours, and then the solids were removed by simple filtration. In the same manner as above, the method of extracting ferulic acid was repeated by adding the added wheat bran and the bacterium enzyme, and the ferulic acid liquid extracted by the second extraction or the tertiary extraction was prepared. Quantity and ferulic acid concentration are shown in Table 1. The pH of the ferulic acid liquid was 6.0 to 6.2, and all of the ferulic acid liquids were used after adjusting the pH to 4.6 by adding citric acid for food. In addition to ferulic acid, the ferulic acid solution contains a small amount of 4-vinyl gua alcohol, which is an intermediate in the process of producing 4-ethyl gua alcohol from ferulic acid, which is 4- It becomes a raw material of ethyl guiacol. Therefore, in the present invention, in order to express the actual raw material of 4-ethyl guiacol as the amount of ferulic acid, the amount of ferulic acid is added to the amount of 4-vinyl guiacol in terms of ferulic acid, which is expressed as the amount of ferulic acid.
(2) 칸디다속 효모의 배양(2) Cultivation of Candida Yeast
효모균으로서는, 발효시험에 의하여, 4-에틸 구아이아콜 생성능이 높은 균주로서, 선발된 NRRL Y-6652균을 사용하였으며, 선정한 균주는 다음과 같이 제조한 국의 당화액에 배양하였다. 먼저 밀가루에 약 25%(w/w)의 물을 혼합하고, 시루식 증자장치에서 수증기로 1시간 증자한 뒤, 아와모리(awamori) 소주제조에 사용하고 있는 흑국균의 변이주(Aspergillus awamorivar.kawachii)를 배양(30℃, 2일간 배양)하여 국(麴)을 만들었다. 이 국에 4배량(w/w)의 물을 가하고, 상법으로 당화하여 국당화액을 만들었는데, 이 국당화액은 pH가 3.6정도 이므로 그대로 사용하였다.As the yeast, by the fermentation test, the selected strain NRRL Y-6652 was used as a strain having high ability to produce 4-ethyl guai alcohol, and the selected strain was cultured in the saccharification solution of the country prepared as follows. First, about 25% (w / w) of water is mixed with the flour, and steamed for 1 hour with steam in a Shiru steamer . Then, aspergillus awamori var. Kawachii is used for making Awamori shochu. ) Was incubated (30 ° C. for 2 days) to make soup. Four times (w / w) of water was added to the soup, and the saccharified solution was made by saccharification by the conventional method. Since the pH of the saccharified solution was about 3.6, it was used as it is.
선정한 칸디다속 효모의 배양에 있어서는, 100㎖용 진탕 플라스크에 넣은50㎖의 배지에, 한천사면배지에 배양한 균체(신선한 균체) 2백금이를 취하여 접종하고, 30℃로 2일간 진탕배양한 뒤, 확대배양하여 대수증식기 후반의 효모균체액을 발효에 사용하였다. 발효액에 옮기기 직전의 배양액 중 칸디다속 효모의 생존균수는 ㎖당 3.9×108개였다.In the culture of the selected Candida yeast, inoculated with 50 ml of medium (fresh cell) cultured in an agar slope medium in a 50 ml medium in a 100 ml shake flask, and incubated at 30 ° C. for 2 days. In the late stage of expansion, the yeast cell solution was used for fermentation. The viable bacterial count of Candida yeast in the culture immediately before transfer to the fermentation broth was 3.9 × 10 8 per ml.
(3) 4-에틸 구아이아콜 발효(3) 4-ethyl guai alcohol fermentation
1ℓ플라스크에 2차 추출한 페룰산액(페룰산 389.2ppm, 쿠말산 16.1ppm) 540㎖를 넣고, 칸디다속 효모(NRRL Y-6652균)의 배양액(생존균수 3.9×108/㎖) 60㎖를 첨가한 뒤, 면전을 하고 발효시켰다. 발효과정 중, 발효액 온도는 30℃로 유지하고, 발효액은 자기교반기(magnetic stirrer)로 교반하였다. 위와 같이 페룰산액에 효모배양액을 첨가·혼합할 때는, 조제 직후의 발효액(시발 발효액)의 페룰산농도는 350.3ppm, 쿠말산농도는 14.5ppm으로 되고, 칸디다속 효모균수는 3.9×107개 /㎖ 으로 된다.(계산상으로)540 ml of ferulic acid solution (389.2 ppm of ferulic acid, 16.1 ppm of cumulic acid) was added to a 1-liter flask, and 60 ml of culture solution (3.9 × 10 8 / ml) of Candida yeast (NRRL Y-6652) was added. After that, face-to-face fermentation. During the fermentation process, the temperature of the fermentation broth was maintained at 30 ° C., and the fermentation broth was stirred with a magnetic stirrer. When adding and mixing the yeast culture solution to the ferulic acid solution as described above, the ferulic acid concentration of the fermentation broth (starting fermentation broth) immediately after preparation is 350.3 ppm, the cumulic acid concentration is 14.5 ppm, and the number of yeast bacteria in the Candida genus is 3.9 × 10 7 / It is in ml. (Calculated)
발효과정 중, 발효액을 취하여 여과하고, 그 여액의 일반성분 및 페놀화합물을 분석한 결과는 표 2 및 표 3과 같다. (페놀화합물은 고속액체크로마토그래피에 의하여 분석함)During the fermentation process, the fermentation broth was taken and filtered, and the results of analyzing the general components and phenolic compounds of the filtrate are shown in Tables 2 and 3. (Phenol compound is analyzed by high performance liquid chromatography)
발효과정에서는, 페룰산에서 4-에틸 구아이아콜이 생성되는 반응이 주로 이루어지는데, 4-에틸 구아이아콜의 생성율은 발효 1.5일 후에 91.5%, 2일 후에는 93.4%로서, 생성율이 양호하였으며, 4-에틸 구아이아콜의 생성은 단기간에 종료되었다.In the fermentation process, 4-ethyl guai alcohol is mainly produced in ferulic acid. The production rate of 4-ethyl guai alcohol was 91.5% after 1.5 days of fermentation and 93.4% after 2 days. The production of 4-ethyl guaicol was terminated in a short time.
발효 중에는 4-에틸 구아이아콜의 생성과 함께, 쿠말산(p-coumaric acid)으로부터 4-에틸 페놀(4-ethyl phenol)이 생성되나 그 양은 매우 적었고 향미 가치도 낮으므로, 여기서는 고려대상으로 하지 않았다. 본 발효법으로 1.5∼2일 발효시킨 발효액은 4-에틸 구아이아콜의 함량이 250ppm 이상이고, 이 양은 발효식품인 간장의 경우에 비하여, 개략 100배 또는 그 이상으로 많은 양이다.During fermentation, 4-ethyl guiacol is produced and 4-ethyl phenol is produced from p-coumaric acid, but the amount is very small and the flavor value is not considered. Did. The fermentation broth fermented 1.5 to 2 days by this fermentation method has a content of 4-ppm guai alcohol of 250 ppm or more, which is approximately 100 times or more than that of soy sauce, which is fermented food.
(실시 예 2) 향미액 사용에 의한 식품의 향미 개선 시험(Example 2) flavor improvement test of food by using flavor liquid
S사 제품인 혼합간장(산분해간장 93%와 양조간장 7%를 혼합한 것)에 (실시 예 1)의 (3)에서 1.5일간 발효시킨 발효액 즉, 향미액(4-에틸 구아이아콜 함량 약 250ppm)을 첨가하되, 4-에틸 구아이아콜로서, 0∼5ppm 범위로 첨가량을 달리한 시험구를 만들고, 6시간 뒤에 5명의 패널에 의하여 관능시험 하였다. ( 향 시험시는 간장원액 시료를 그대로 사용하고, 맛 시험시는 간장원액 시료를 물로 5배 희석한 것을 사용하였다.) 시험결과, 향미액을 적당량 첨가한 것은 무첨가의 것(대조구)에 비하여, 산분해 간장에 기인하는 불쾌취가 탈취되고(masking작용에 의함), 양조 간장의 특징적인 향이 부가되어, 간장향이 현저하게 개선되었으며, 간장의 짠맛은 완화되어 부드러워지고 맛도 개선되었다고, 패널전원이 동의하였다. 관능상으로는 간장에 4-에틸 구아이아콜로서, 2ppm정도 첨가한 것이 적당하다고 하였다.4-에틸 구아이아콜을 주성분으로 하는 향미액(향미료)을 식품의 향미개선에 사용하는 방법은 본 실시예 외에 각종 조미액, 간장, 된장, 주류 등의 제조에서도 유용한 재료로 사용될 수 있다. 또 훈연식품 제조시 훈증액으로 사용될 수도 있다.Fermentation broth fermented for 1.5 days in (3) of (Example 1) to mixed soy (mixed soy sauce 93% and brewed soy sauce) manufactured by S Company, that is, flavoring liquid (4-ethyl guiacol content 250ppm) was added, and 4-ethyl guacol was used to make test pieces with different amounts of addition in the range of 0 to 5 ppm, and 6 hours later, the sensory test was carried out by five panels. (In the flavor test, the soy sauce sample was used as it was, and in the taste test, the soy sauce sample was diluted 5 times with water.) As a result of the test, an appropriate amount of flavor solution was added as compared to the no addition (control). The unpleasant odor caused by acid-decomposed soy sauce was deodorized (by masking action), the characteristic flavor of brewed soy sauce was added, so that the soy sauce flavor was remarkably improved, and the salty taste of soy was softened and the taste was improved. Agreed. Functionally, it is said that it is appropriate to add about 2 ppm of 4-ethyl guai alcohol to soy sauce. The method of using a 4-ethyl guia alcohol-based flavoring liquid (flavor) for improving the flavor of food is described in this example. It can be used as a useful material in the manufacture of various seasoning liquids, soy sauce, miso, and liquor. It can also be used as a fumigation liquid in the manufacture of smoked food.
본 발명의 제조법에 의하여, 페룰산(ferulic acid)원료에서, 4-에틸 구아이아콜(4-ethyl guaiacol)의 함량이 높은 향미액(향미료)를 단기간에, 효율적으로 제조하는 것이 가능하다. 또 4-에틸 구아이아콜 생성능이 높은 칸디다속 효모균주는 4-에틸 구아이아콜 함량이 높은 향미액(향미료)의 제조에 유효하게 사용할 수 있다.According to the production method of the present invention, it is possible to efficiently produce a flavor liquid (flavour) having a high content of 4-ethyl guaiacol from ferulic acid raw materials in a short time. Candida yeast strain with high 4-ethyl guai alcohol production ability can be effectively used for the preparation of flavor liquid (flavor) with high 4-ethyl guai alcohol content.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0885969A1 (en) * | 1997-06-13 | 1998-12-23 | Lutz Dr. Laurisch | Culture medium to detect Streptococcus mutans |
| JP2000125840A (en) * | 1998-10-29 | 2000-05-09 | Kikkoman Corp | Production of shochu |
| KR100280270B1 (en) * | 1998-09-29 | 2001-02-01 | 박윤중 | Food flavor improvement method |
| US6235507B1 (en) * | 1997-06-19 | 2001-05-22 | Givaudan Roure (International) Sa | Microbiological process for producing vanillin |
| KR20010095935A (en) * | 2000-04-12 | 2001-11-07 | 박윤중 | Femented liquor for flavoring |
-
2001
- 2001-07-11 KR KR10-2001-0041775A patent/KR100460605B1/en not_active IP Right Cessation
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0885969A1 (en) * | 1997-06-13 | 1998-12-23 | Lutz Dr. Laurisch | Culture medium to detect Streptococcus mutans |
| US6235507B1 (en) * | 1997-06-19 | 2001-05-22 | Givaudan Roure (International) Sa | Microbiological process for producing vanillin |
| KR100280270B1 (en) * | 1998-09-29 | 2001-02-01 | 박윤중 | Food flavor improvement method |
| JP2000125840A (en) * | 1998-10-29 | 2000-05-09 | Kikkoman Corp | Production of shochu |
| KR20010095935A (en) * | 2000-04-12 | 2001-11-07 | 박윤중 | Femented liquor for flavoring |
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| KR20030006182A (en) | 2003-01-23 |
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