In order to improve taste quality, enzymatic modification of stevia by grafting glucose was investigated using soluble starch as glucose donor. The hydrolysis was catalyzed by acid-resistant mesophilic α-amylase, whose trade name is Taka-diastase, that was screened from three different α-amylases. Using stevioside conversion rate as an indicator, the enzymatic processes were optimized and the optimum conditions obtained were donor starch concentration 100 g/L, substrate ratio 1:10 (m/m), reaction temperature 60 ℃, and enzyme concentration 1.50 U/mL. The conversion rate of the modified stevioside was determined as 42.5% by high performance liquid chromatography (HPLC). The organoleptic evaluation results showed that the enzymatic modification improved the taste quality of stevia significantly. The sweetness became mild and soft, and a slight bitter taste still remains at high concentration, but disappears at low concentration. Desirable taste quality with significantly reduced production cost could be achieved by enzymatic modification of stevia.
[1] 娄力行.甜菊糖及其衍生物的研究进展[J].中国糖料,2008,2: 70-72
LOU Li-xing. Research progress of stevioside and steviol [J]. Sugar Crops of China, 2008, 2: 70-72
[2] Jentzer J B, Alignan M, Vacagarcia C, et al. Response surface methodology to optimise accelerated solvent extraction of steviol glycosides from stevia rebaudiana Bertoni leaves [J]. Food Chemistry, 2015, 166: 561-567
[3] Genus J M C. Molecules of interest stevioside [J]. Phytochemistry, 2003, 64(5): 913-921
[4] Prakash I, Dubois G E, Clos J F, et al. Development of rebiana, a natural, non-caloric sweetener [J]. Food & Chemical Toxicology, 2008, 46(7): S75-S82
[5] 李培.甜菊糖的重结晶分离及酶法改性[D].无锡:江南大学, 2009
LI Pei. Recrystallization separation and enzymatic modification of stevioside [D]. Wuxi: Jiangnan University, 2009
[6] 万会达,蔡亚,校秋燕,等.甜菊糖的酶法改性及其生物活性研究进展[J].中国食品添加剂,2011,5:188-196
WAN Hui-da, CAI Ya, XIAO Qiu-yan, et al. Recent advance in steviol glucosides modification and bioactivities [J]. China Food Additives, 2011, 5: 188-196
[7] Suzuki K, Fukumura T, Shibasaki-Kitakawa N, et al. Kinetic model for synthesis of fructosyl-stevioside using suspended β-fructofuranosidase [J]. Biochemical Engineering Journal, 2002, 10(3): 207-215
[8] Pól J, Ostrá E V, Karásek P, et al. Comparison of two different solvents employed for pressurised fluid extraction of stevioside from stevia rebaudiana : methanol versus water [J]. Analytical & Bioanalytical Chemistry, 2007, 388(8): 1847- 1857
[9] 王贵民,董振红,郝再彬.甜叶菊糖苷的应用和安全性的研究进展[J].中国食品添加剂,2007,6:65-69
WANG Gui-ming, DONG ZHEN-hong, HAO Zai-bin. Application and safety of steviol glycoside in Ieaves of stevia rebaudiana (bertoni) bertoni [J]. China Food Additives, 2007, 6: 65-69
[10] Kaneda N, Kasai R, Yamasaki K, et al. Chemical studies on sweet diterpene-glycosides of stevia rebaudiana: Conversion of stevioside into rebaudioside-A [J]. Chemical & Pharmaceutical Bulletin, 1977, 25(9): 2466-2467
[11] Kitahata S, Ishikawa H, Miyata T, et al. Production of rubusoside derivatives by transgalactosylation of various α-galactosidases (Biological Chemistry) [J]. Journal of the Agricultural Chemical Society of Japan, 1989, 53(11): 2923-2928
[12] 叶发银,杨瑞金,华霄,等.中温α-淀粉酶改性甜菊糖的研究[J].食品与发酵工业,2012,38(6):57-60
YE Fa-yin, YANG Rui-jin, HUA Xiao, et al. Enzymatic modification of steviol glycosides by medium-temperature α-amylase [J]. Food Science & Technology, 2012, 38(6): 57-60
[13] 徐仲伟,李娜,宁正祥,等.酶法改质甜菊糖的研究[J].食品科技,2008,33(12):35-39
XU Zhong-wei, LI Na, NING Zheng-xiang, et al. Study on enzymatic modification of stevioside [J]. Food Science & Technology, 2008, 33(12): 35-39
[14] 赵露,孙颖.甜菊糖酶法改性的工艺研究[J].粮食与食品工业,2013,20(4):67-70
ZHAO Lu, SUN Ying. Technical research on enzymatic modification of stevia sugar [J]. Cereal & Food Industry, 2013, 20(4): 67-70
[15] Zimmermann B F. Tandem mass spectrometric fragmentation patterns of known and new steviol glycosides with structure proposals [J]. Rapid Communications in Mass Spectrometry, 2011, 25(11): 1575-1582
[16] Kochikian V T, Markosian A A, Abelian L A, et al. Combined enzymatic modification of stevioside and rebaudioside A [J]. Applied Biochemistry & Microbiology, 2006, 42(1): 31-37
[17] 王德骥.关于甜菊糖苷的甜度,甜味和苦涩后味的成因机理[J].中国食品添加剂,2007,3:46-53
WANG De-ji. Sweetness and mechanism of picric in stevioside [J]. China Food Additives, 2007, 3: 46-53
[18] 叶发银.高纯度莱鲍迪苷A的制备和甜菊苷的酶法改性研究[D].江南大学,2013
YE Fa-yin. Preparation of high-purity rebaudioside a and enzymatic modification of stevioside [D]. Jiangnan University, 2013
[19] 徐仲伟,邵佩霞,陈华勇,等.β-呋喃果糖苷酶催化甜菊糖的改性研究[J].现代食品科技,2008,24(11):1108-1110
XU Zhong-wei, SHAO Pei-xia, CHEN Hua-yong, et al. Optimization of enzymatic modification of steviosides [J]. Modern Food Science & Technology, 2008, 24(11): 1108-1110