TW201629208A - 維克哈默酵母菌（Ｗｉｃｋｅｒｈａｍｏｍｙｃｅｓ）屬微生物培養物 - Google Patents
維克哈默酵母菌（Ｗｉｃｋｅｒｈａｍｏｍｙｃｅｓ）屬微生物培養物 Download PDFInfo
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Abstract
本發明提供一種大量含有苯甲酸乙酯，具有較化學合成品更複雜且更新鮮之如水果般之香味的培養物。
一種培養物，其係將屬於維克哈默酵母菌屬之微生物於含乳成分之培養基中進行培養而獲得。
Description
本發明係關於一種具有良好香氣之培養物及其用途。
先前已知有，使用各種產生香氣成分之酵母、例如釀酒酵母等，而獲得具有良好香氣之培養物的方法，該培養物係用作飲食品(專利文獻1~3)。
作為香氣成分之一的苯甲酸乙酯於化學上可由苯甲酸與乙醇合成。該化合物已知為楊桃之香氣成分之一，於產業上用作果實香味之人工香料。亦已知有使用含有苯甲酸乙酯作為香氣成分之釀酒酵母的培養物，其含量較少而為0.006~0.009ppm(非專利文獻1)。
又，皮傑普維克哈默酵母菌(Wickerhamomyces pijperi)係自皮傑普畢赤酵母(Pichia pijperi)再分類之一種酵母，非專利文獻2中記載有將該酵母於酵母萃取物-甘露醇(yeast extract-mannitol)培養基、酵母-蛋白腖-葡萄糖(yeast-peptone-dextrose，YPD)培養基中進行培養，所獲得之培養物之特性不明，對於可於含乳成分之培養基中進行培養及其培養物之香氣完全未知。
[專利文獻1]日本專利特開平5-49465號公報
[專利文獻2]日本專利特開2001-103959號公報
[專利文獻3]日本專利特開2012-55286號公報
[非專利文獻1]Journal of the institute of Brewing, 116, 190-196, 2010
[非專利文獻2]Microbiology, 155, 3142-3148, 2009
上述苯甲酸乙酯等香氣成分之化學合成品之香氣單調，作為香料用組合物而言未必令人滿意。因此，業界謀求一種能夠利用於化妝品或飲食品、醫藥品，具有嗜好性優於化學合成品之香味，且能夠用作香料用組合物之組合物。又，嗜好如水果般之香味之消費者亦較多，謀求提供如水果般之新穎香味。
本發明之課題在於提供一種組合物及其用途，該組合物係能夠利用於香料用組合物、化妝品、飲食品及醫藥品等之新穎組合物，並且大量含有苯甲酸乙酯，具有較苯甲酸乙酯之化學合成品更複雜且更新鮮之如水果般之香味。
因此，本發明者為了開發出具有良好香氣之組合物而進行了各種研究，結果完全意外地發現，於將維克哈默酵母菌屬之微生物於含有乳成分之培養基中進行培養時，獲得大量含有作為香氣成分已知之苯甲酸乙酯，且具有較苯甲酸乙酯之化學合成品更複雜且更新鮮之如水果般之香氣的培養物，該培養物作為可用於香料用組合物、化妝品、飲食品、醫藥品等之培養物有用，從而完成本發明。
即，本發明提供以下之[1]至[10]。
[1]一種培養物，其係將屬於維克哈默酵母菌屬之微生物於含乳成分之培養基中進行培養而獲得。
[2]如[1]之培養物，其中含乳成分之培養基為含乳清之培養基。
[3]如[1]或[2]之培養物，其中屬於維克哈默酵母菌屬之微生物為皮傑普維克哈默酵母菌。
[4]如[1]至[3]中任一項之培養物，其中屬於維克哈默酵母菌屬之微生物為皮傑普維克哈默酵母菌NBRC1290及/或皮傑普維克哈默酵母菌NBRC1887。
[5]如[1]至[4]中任一項之培養物，其中含乳成分之培養基係進而含有苯甲酸及/或其鹽者。
[6]一種香料用組合物，其含有如[1]至[5]中任一項之培養物。
[7]一種化妝品，其含有如[1]至[5]中任一項之培養物。
[8]一種飲食品，其含有如[1]至[5]中任一項之培養物。
[9]一種醫藥品，其含有如[1]至[5]中任一項之培養物。
[10]一種對化妝品、飲食品或醫藥品賦予香味之方法，其特徵在於：調配如[1]至[5]中任一項之培養物。
本發明之培養物由於大量含有苯甲酸乙酯，且具有較苯甲酸乙酯之化學合成品更複雜且更新鮮之如水果般之香味，故而不僅作為香料用組合物或化妝品有用，亦作為飲食品或醫藥品所使用之培養物有用。
本發明之培養物係將屬於維克哈默酵母菌屬之微生物於含乳成分之培養基中進行培養而獲得之培養物。
作為屬於維克哈默酵母菌屬之微生物，可列舉：皮傑普維克哈默酵母菌(Wickerhamomyces pijperi)、異常維克哈默酵母菌(Wickerhamomyces anomalus)、牛維克哈默酵母菌(Wickerhamomyces
bovis)、臘寶維克哈默酵母菌(Wickerhamomyces rabaulensis)、漢普夏維克哈默酵母菌(Wickerhamomyces hampshirensis)、斯特拉斯堡維克哈默酵母菌(Wickerhamomyces strasburgensis)、賽道威維克哈默酵母菌(Wickerhamomyces sydowiorum)、Wickerhamomyces lynferdii、西弗維克哈默酵母菌(Wickerhamomyces ciferrii)、錢巴德維克哈默酵母菌(Wickerhamomyces chambardii)、森林維克哈默酵母菌(Wickerhamomyces silvicola)、雙孢維克哈默酵母菌(Wickerhamomyces bisporus)、赤楊維克哈默酵母菌(Wickerhamomyces alni)、加拿大維克哈默酵母菌(Wickerhamomyces canadensis)、指甲維克哈默酵母菌(Wickerhamomyces onychis)、土壤維克哈默酵母菌(Wickerhamomyces edaphicus)、擲孢維克哈默酵母菌(Wickerhamomyces patagonicus)等。其中，更佳為皮傑普維克哈默酵母菌。進而，皮傑普維克哈默酵母菌中，進而較佳為寄存於NBRC(NITE Biological Resource Center，日本技術評價研究所生物資源中心)之NBRC1290、NBRC1887。再者，該等微生物可單獨使用，亦可將複數種組合使用。再者，於舊分類中分類為皮傑普畢赤酵母(Pichia pijperi)且新分類為皮傑普維克哈默酵母菌者，包括在本申請案之皮傑普維克哈默酵母菌中。
於本說明書中，所謂「乳成分」係指含有牛乳、山羊乳、綿羊乳等獸乳之生乳、加熱乳、脫脂粉乳、全脂粉乳或者生奶油、乳清等源自乳之成分者。
作為含乳成分之培養基，只要為含有乳成分之培養基，則亦可進而含有其他成分。作為該其他成分，並無特別限定，較佳為葡萄糖、半乳糖、乳糖、果糖等糖，苯甲酸及/或其鹽。作為糖，就被微生物同化之方面而言，較佳為葡萄糖。又，就添加後提高微生物之苯甲酸乙酯之產生量之方面而言，較佳為苯甲酸及/或其鹽，作為苯甲
酸之鹽，更佳為苯甲酸鈉及苯甲酸鉀等苯甲酸鹼金屬鹽，進而較佳為苯甲酸鈉。因此，作為苯甲酸及/或其鹽，較佳為添加苯甲酸及/或苯甲酸鈉。
作為乳成分，乳清由於會提高微生物之苯甲酸乙酯之產生量，故而尤佳。所謂「乳清」係指自牛乳去除乳脂肪分、酪蛋白而獲得者。例如，乳清可以將乳成分利用微生物進行醱酵時之培養上清液之形式獲得，較佳為含有乳糖、半乳糖等糖、胺基酸、乳酸、蛋白質等者。又，作為乳清，尤佳為將乳成分利用乳酸菌及/或雙歧桿菌進行培養而獲得之培養上清液。此處，作為乳酸菌及/或雙歧桿菌，可列舉：酪蛋白乳酸桿菌、馬里乳酸桿菌、嗜酸乳酸桿菌、德氏乳酸桿菌德氏亞種、瑞士乳酸桿菌等乳酸桿菌屬細菌，嗜熱鏈球菌等鏈球菌屬細菌，雷特氏乳球菌雷特氏亞種、雷特氏乳球菌乳脂亞種等乳球菌屬細菌，糞腸球菌等腸球菌屬細菌，或者短型雙叉桿菌、雙叉雙叉桿菌、龍根雙叉桿菌等雙叉桿菌屬細菌。
培養基中之乳成分之含量並無特別限定，以固形物成分換算，較佳為1~50質量%(以下，簡稱為「%」)，更佳為2~10%。又，就微生物之苯甲酸乙酯之產生量及微生物之增殖能力之方面而言，苯甲酸及/或其鹽之含量於含乳成分之培養基中較佳為0.001~0.1%，更佳為0.01~0.05%。進而，就微生物之增殖能力之方面而言，葡萄糖之含量較佳為0.1~10%，更佳為1~5%。
屬於維克哈默酵母菌屬之微生物之培養溫度並無特別限定，就培養物之香味之方面而言，較佳為15~30℃，進而較佳為20~30℃。培養時間較佳為8小時以上，進而較佳為24~32小時。又，於將苯甲酸及/或其鹽添加至培養基中之情形時，較佳為24~48小時，進而較佳為32~48小時。作為培養方法，可列舉：攪拌培養、靜置培養、振盪培養、中和培養等。
本發明之培養物中包含1.0ppm以上之苯甲酸乙酯，具有較苯甲酸乙酯之化學合成品更複雜且更新鮮之如水果般之香氣。苯甲酸乙酯之含量較佳為1.0~50ppm，更佳為1.0~30ppm。
又，本發明之培養物中，亦可包含苯甲酸乙酯以外之其他香氣成分。作為其他香氣成分，可列舉異戊醇等香氣成分。
本發明之培養物由於大量含有苯甲酸乙酯，具有且較苯甲酸乙酯之化學合成品更複雜且更新鮮之如水果般之良好香氣，故而可利用於香料用組合物、化妝品、飲食品及醫藥品等。於化妝品領域，由於工業上可利用之源自生物之香料較少，天然香料之需求高於合成香料，故而可適宜地利用於化妝品。
對於本發明之培養物，亦可將培養物直接調配至香料用組合物、化妝品、飲食品及醫藥品等中，較佳為調配經過濾者。又，亦可藉由冷凍乾燥法或者蒸餾法等對培養物進行濃縮，並將該濃縮物調配至香料用組合物、化妝品、飲食品及醫藥品等中。
作為化妝品，可列舉：香水、古龍水、淡香水等芳香化妝品，化妝水、乳液、化妝水、乳霜、面膜、美容液等基礎化妝品，洗髮精或潤絲精等頭髪用化妝品，入浴劑等浴用化妝品、粉底等化妝用化妝料、防曬霜等特殊化妝品等。作為飲食品，可列舉：各種清涼飲料、發泡酒、啤酒、清酒、點心類、冰點心類、霜淇淋、醱酵乳等乳製品等。又，作為醫藥品，可列舉：乳霜、軟膏、凝膠等外用劑等。該等化妝品、飲食品、醫藥品中之本發明培養物之含量以固形物成分換算較佳為0.01~10%，更佳為0.1~1%。
[實施例]
其次，列舉實施例詳細地說明本發明。
實施例1
(1)菌株及前培養
(i)使用菌株
皮傑普維克哈默酵母菌YIT8095(NBRC 1290)、皮傑普維克哈默酵母菌YIT12779(NBRC 1887)
比較例：馬克斯克魯維酵母(Kluyveromyces marxianus)YIT12612(NBRC 0260)
(ii)前培養
前培養：向2ml之酵母及黴菌(Yeast and Mold，YM)用培養基(1%葡萄糖、0.5%蛋白腖、0.3%酵母萃取物、0.3%麥芽萃取物)中接種20%甘油冷凍保存株20μL，於30℃、160rpm下振盪培養24小時。
(2)菌所產生之香氣成分之分析
(i)前培養
前培養：向包含1%葡萄糖之2ml之含乳清之培養基中接種前培養中所獲得之培養液20μL，於30℃、160rpm下振盪培養24小時。
作為上述含乳清之培養基，使用將嗜熱鏈球菌(Streptococcus thermophilus)YIT2084(FERMBP-10879)於3%脫脂粉乳培養基中進行培養而獲得之培養物之上清液。對該上清液進行分析，結果包含1.1%之乳糖、0.4%之半乳糖、0.4%之乳酸，pH值為4.0。
(ii)培養
培養：向包含1%葡萄糖之5ml之含乳清之培養基中接種前培養液50μL，於30℃、160rpm下振盪培養24小時。
(iii)香氣成分之分析
培養完畢後，試樣係於8000rpm下離心分離5分鐘，取2ml之上清液置於20ml之小瓶中，並藉由頂隙氣相層析質譜法(Headspace gas chromatography mass spectrometry，HS-GC-MS)鑑定培養物中之香氣成分(表1、表2)。又，所鑑定之香氣成分之定量係利用HS-GC-火焰游離偵檢器(Flame Ionization Detector，FID)而進行(表3、表4)。將結果
示於表5。又，基於下述基準進行培養液之香味之官能檢査。將其結果示於表5。
(官能檢査之評價基準)
5：強烈地發出複雜且新鮮之如水果般之香味。
4：發出複雜且新鮮之如水果般之香味。
3：略微發出複雜且新鮮之如水果般之香味。
2：幾乎不發出略微發出複雜且新鮮之如水果般之香味。
1：不發出複雜且新鮮之如水果般之香味。
自NBRC1290株與NBRC1887株之培養液強烈地感覺到複雜且新鮮之如水果般之香味，對香氣成分進行分析，並將各化合物之含量加以比較，結果特徵為苯甲酸乙酯之含量較多。作為與該等培養液之比較，對將另一種酵母(Kluyveromyces marxianus NBRC0260株)於含乳
清之培養基中進行培養而獲得之培養液之香氣成分進行分析、評價，結果如薔薇般之香味強烈，但幾乎不發出如水果般之香味，不含有苯甲酸乙酯。
進行NBRC1290株及NBRC1887株之培養液、及與培養液中等量之苯甲酸乙酯之化學合成品單獨之香味之官能檢査，結果對於NBRC1290株及NBRC1887株之培養液，感覺到化學合成品不會發出之香味之複雜感，而為優於化學合成品之香味。
(3)根據培養基之菌之香氣成分產生量之差異
使用NBRC1290株及NBRC1887株進行實驗。
(i)前培養
前培養：向包含1%葡萄糖之2ml之含乳清之培養基、2ml之YPD培養基(1%酵母萃取物、2%蛋白腖、2%葡萄糖)、2ml之YM培養基(1%葡萄糖、0.5%蛋白腖、0.3%酵母萃取物、0.3%麥芽萃取物)中，分別接種前培養中所獲得之培養液20μL，於30℃、160rpm下振盪培養24小時。
(ii)培養
培養：向包含1%葡萄糖之2ml之含乳清之培養基、2ml之YPD培養基、2ml之YM培養基中，分別接種前培養中所獲得之培養液5ml，於30℃、160rpm下振盪培養24小時。
(iii)香氣成分之含量之分析
培養完畢後，利用HS-GC-FID對培養物中之香氣成分之含量進行分析(表3、表4)。將結果示於表6。又，以與實施例1(2)相同之基準進行培養物之香味之官能檢査。將其結果示於表6。
自於含乳清之培養基中培養NBRC1290株與NBRC1887株而獲得之培養液強烈地感覺到複雜且新鮮之如水果般之香味，培養液大量含有苯甲酸乙酯。另一方面，有時於YM培養基、YPD培養基中培養上述菌株而獲得之培養液中亦含有苯甲酸乙酯，但其含量較少，幾乎不發出複雜且新鮮之如水果般之香味。
(4)根據培養條件之菌之香氣成分產生量之差異
使用NBRC1887株進行實驗。
(i)前培養
前培養：向包含1%葡萄糖之2ml之含乳清之培養基中接種前培養中所獲得之培養液20μL，於30℃、160rpm下振盪培養24小時。
(ii)培養
培養：向包含1%葡萄糖之100ml之含乳清之培養基中接種前培養液1ml，於15℃、20℃、25℃、30℃之培養條件下，於160rpm下進行振盪培養。又，於培養0、24、32、48、56小時之時，回收樣品。
(iii)苯甲酸乙酯之含量之分析
利用HS-GC-FID對各培養物中之苯甲酸乙酯之含量進行分析(表3、表4)。將結果示於表7。又，基於下述基準進行培養物之香味之官能檢査。將其結果示於表7。
(官能檢査之評價基準)
◎：強烈地發出複雜且新鮮之如水果般之香味。
○：發出複雜且新鮮之如水果般之香味。
△：略微發出複雜且新鮮之如水果般之香味，但複雜感較弱。
▲：其他香氣成分之香味較強，香味之特徵發生變化，發出如日本酒般之香味。
×：不發出複雜且新鮮之如水果般之香味。
於培養溫度為15℃之情形時，雖然藉由微生物之醱酵而產生苯甲酸乙酯，但其他香氣成分之香味較弱，培養物之香味之複雜感較弱。另一方面，於20~30℃下進行培養而獲得之培養物發出複雜且新鮮之如水果般之香味，香味良好。又，若培養時間達到48小時以上，則因微生物之醱酵，苯甲酸乙酯以外之香氣成分之產生量會上升，因此其他香氣成分之香味變強，香味之特徵發生變化，發出如日本酒般之香味。因此，作為培養時間，宜為24~32小時。
(5)將苯甲酸或其鹽調配至培養基中時之苯甲酸乙酯含量之差異使用NBRC1887株進行實驗。
(i)前培養
前培養：向包含1%葡萄糖之2ml之含乳清之培養基中接種前培養中所獲得之培養液20μl，於30℃、160rpm下振盪培養24小時。
(ii)培養
培養：向包含1%葡萄糖之100ml之含乳清之培養基中，以成為0.01%之方式調配苯甲酸鈉。又，同樣地，向包含1%葡萄糖之100ml之含乳清之培養基中，以成為0.01%之方式調配苯甲酸，以成為0.01%、0.05%、0.1%之方式調配苯丙胺酸，以成為0.01%、0.05%之方式調配對羥基甲基苯。進而，作為對照，亦準備不調配添加物之包含1%葡萄糖之100ml之含乳清之培養基。向各培養基中接種前培養液1ml，於30℃、160rpm下進行振盪培養。又，於培養0、24、32、48小時之時，回收樣品。
(iii)苯甲酸乙酯之含量之分析
利用HS-GC-FID對各培養物中之苯甲酸乙酯之含量進行分析(表3、表4)。將結果示於表8。
於調配有苯甲酸或苯甲酸鈉之情形時，於培養32~48小時之時，培養物中之苯甲酸乙酯之含量顯著增加。於調配有其他物質之情形時，培養物中之苯甲酸乙酯之含量幾乎未增加。
製造例1
以表9之組成將(5)與(6)混合，向其中添加(1)~(4)並充分地攪拌，而製備化妝水。該化妝水發出複雜且新鮮之如水果般之香味。再者，作為培養液，使用如下者，即，向包含1%葡萄糖之含乳清之培養基100ml中接種實施例1(4)之前培養中所獲得之前培養液1ml，於
20℃、24小時、160rpm下進行振盪培養而獲得培養液，對該培養液進行過濾而獲得者。
製造例2
以表10之組成，對(10)添加(7)~(9)並加溫，於80℃下添加(1)~(6)並進行乳化，冷卻至室溫，而製備乳液。該乳液發出複雜且新鮮之如水果般之香味。再者，培養液係使用與製造例1相同者。
製造例3
以表11之組成，對(12)添加(9)~(11)並加溫，於80℃下添加(1)~(8)並進行乳化，冷卻至室溫，而製備乳霜。該乳霜發出複雜且新鮮之如水果般之香味。再者，培養液係使用與製造例1相同者。
Claims (10)
- 一種培養物，其係將屬於維克哈默酵母菌屬之微生物於含乳成分之培養基中進行培養而獲得。
- 如請求項1之培養物，其中含乳成分之培養基為含乳清之培養基。
- 如請求項1之培養物，其中屬於維克哈默酵母菌屬之微生物為皮傑普維克哈默酵母菌。
- 如請求項1之培養物，其中屬於維克哈默酵母菌屬之微生物為皮傑普維克哈默酵母菌NBRC1290及/或皮傑普維克哈默酵母菌NBRC1887。
- 如請求項1之培養物，其中含乳成分之培養基係進而含有苯甲酸及/或其鹽者。
- 一種香料用組合物，其含有如請求項1至5中任一項之培養物。
- 一種化妝品，其含有如請求項1至5中任一項之培養物。
- 一種飲食品，其含有如請求項1至5中任一項之培養物。
- 一種醫藥品，其含有如請求項1至5中任一項之培養物。
- 一種對化妝品、飲食品或醫藥品賦予香味之方法，其特徵在於：調配如請求項1至5中任一項之培養物。
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| CN111278326A (zh) | 2017-07-28 | 2020-06-12 | 轨迹Ip有限责任公司 | 改善皮肤、头发和头皮健康的基于酵母的面膜 |
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