采用微生物发酵制取丙酮酸的研究
摘要
采用微生物发酵技术制取丙酮酸，利用物理的和化学的方法进行诱变，获得一株汉逊德巴利酵母汉逊变种NO39-1。通过优化培养条件，该突变菌株生产丙酮酸的能力较出发菌株提高了1.8倍。在30℃，200r／min摇瓶培养72h的条件下，发酵液中丙酮酸含量达到5.2g/L。
Fermentative production of pyruvic acid by Debaryomyces hansii var. hansenii was investigated in this paper. A mutant was obtained after DES and UV treatment. By using orthogonal experiment method, the concentration of pyruvic acid reached 5.2g/L at 30℃,200r/min,72h in flask culture.
Fermentative production of pyruvic acid by Debaryomyces hansii var. hansenii was investigated in this paper. A mutant was obtained after DES and UV treatment. By using orthogonal experiment method, the concentration of pyruvic acid reached 5.2g/L at 30℃,200r/min,72h in flask culture.
引文
1 Robert C. Weast, Melvin J. Astle. CRC Handbook of Chemistry and Physics. 63~(rd).1982-1983
2 段长强，孟庆芳，张泰等．现代化学试剂手册(第一分册)[M]．北京：化学工业出版社，1988
3 潘竹林．丙酮酸缺陷症研究现状．中华血液学杂志．1999，20(4)：223-224
4 唐胜南，李艳，伍期专．血乳糖、丙酮酸比值测定的意义．中国卫生检验杂志．1999，9(6)：455-456
5 谭宁宁，王荣珍，曹维娟．呼吸道感染患儿血液丙酮酸含量测定及临床观察．江苏医药．1996，22(2)：116
6 Yonehara T. Miyata R. Fermentative production of pyruvate from glucose by Torulgpsis glabrata. J Ferment Bioeng. 1994,78:155-159
7 Nagasawa T, Utagawa T, Goto J, Kim CJ, Tani Y, Kumagai H, Yamada H. Synthesis of L-tyrosine-related amino acids by tyrosine phenol-lyase of Citrobacter intermedius Eur J Biochem.1981, 117:33-40
8 姜胜斌，廖桂英，袁华．丙酮酸系列产品．精细与专用化学品．2000，21：8-9
9 袁辉，华子春．丙酮酸发酵能力的提高．微生物学杂志．2000，20(3)：23-24，39
10 Stanko, R.T., Robertson, R.J., Spina, R.J., ReAlly, J.J., Greenawalt, K.D., Goss, F.L. Enhancement of arm-exercise endurance capacity with dihydroxyacetone and pyruvate. Journal of Applied Physiology. 1990,68:119-124
11 Stanko, R.T., Robertson, R.J., Galbreath, R.W., ReAlly, J.J., Greenawalt, K.D., Goss, F.L. Enhanced leg-exercise endurance with a high-carbohydrate diet and dihydroxyacetone and pyruvate. Journal of Applied Physiology. 1990,69:1651-1656
12 Robertson, KJ., Stanko, R.T., ReAlly, J.J., Greenawalt, K.D., Goss, F.L., Spina, R.J. Blood glucose extraction as a mediator of perceived exertion during prolonged exercise. European Journal of Applied Physiology. 1990,61:100-105
13 Stanko, RT. Tietze, D.L., Arch, J.E. Body composition, energy utilization, and nitrogen metabolism with a 4.25-MJ/d low-energy diet supplemented with pyruvate. American Journal of Clinical Nutrition. 1992,56:630-635
14 Stanko, R.T. Tietze, D.L., Arch, i.E. Body composition, energy utilization and nitrogen metabolism with a severely restricted diet supplemented with dihydroxyacetone and pyruvate. American Journal of Clinical Nutrition. 1992,55:771-775
15 Stanko, R.T., Adibi, S.A. Inhibition of lipid accumulation and enhancement of energy expenditure by the addition of pyruvate and dihydroxyacetone to a rat diet. Metabolism. 1986,35:182-186
16 Stanko, R.T., Ferguson, T.L., Newman, C.W., Newman, R.K. Reduction of carcass fat in swine with dietary addition of dihydroxyacetone and pyruvate. Journal of Animal Science. 1989,6:1272-1278
17 Cortez, M.Y., Torgan, C.E., Brozinick, J.T., Miller, R.H., Ivy, J.L. Effects of pyruvate and dihydroxyacetone consumption on the growth and metabolic state of obese Zucker rats. American Journal of Clinical Nutrition. 1991,53:847-853
18 James B. Roufs, M.S., Pyruvate—Does It Amp Endurance and Burn More Fat?", Muscle & Fitness Magazine. Dec. 1996
19 Andy Calhoun, Pyruvate: The Universal Supplement, The Athlete's Power Source Quarterly Winter 97-98
20 吕咏梅．丙酮酸钙及原料丙酮酸发展前景广阔．中国医药情报．2002，8(3)：52-53
21 张庆勇．丙酮酸及丙酮酸钠的制备．化学试剂．1997，19(6)：375，336
22 精细化学产品目录[M]．化学试剂．北京：化学工业出版社．2000
23 傅月华，余炳辉，纪佩芳．应用丙酮酸钠检出食品中亚致死菌的研究．12-14
24 Howard J W, Fraser W A. Preparation of pyruvic acid. Org Synth Coll.1932,1:475-480
25 US 4 136 111,1979
26 JP 7 981 221,1979
27 JP 9 085 230,1990
28 JP 7 939 016,1979
29 吴俊明．1，2-丙二醇空气氧化合成丙酮醛[J]．化工时刊．1997，11(12)：26-28
30 朱京．丙二醇空气氧化制丙酮醛的电结晶银催化剂[J]．复且大学学报．1989，28(1)：14-18
31 Get 3 219 355, 1983
32 Baizer M M. [J]. Tetrahedron. 1984, 40(6): 935-969
33 Chaussard J, Folest J C, et al [J]. Synthesis. 1990,5:369-381
34 廖桂英，姜肚斌，袁华．丙酮酸的电化学合成法．天津化工．2000，6：29
35 UK 2 018 773. 1979
36 J Mol. Catal [J].1992, 71(1):25-35
37 JP 5 619 854,1981
38 JP 5 724 336, 1982
39 Mamoru Ai,.Kyoji Ohdan. Chem Lett[J].1995, (5): 405
40 Mamoru Ai, Kyoji Ohdan. APPL. Catal A. 1997,150(1): 13-20
41 USP 4 229 590, 1980
42 JP 8 697 247,1986
43 胡旭灿等．乳酸乙脂在Ag／NaY催化剂上的脱氢反应制丙酮酸乙酯[J]．催化学报．1998，19(5)：428-431
44 杨辉琼，易翔．乳酸氧气氧化法制备丙酮酸．化学世界．2002，43(6)：307-309
45 杨辉琼，郭贤烙，易翔．双氧水催化氧化乳酸合成丙酮酸．化学工程师．2002，2：14-15
46 廖桂英，王应席．以磷酸铁盐为催化剂：乳酸氧化脱氢制丙酮酸．荆州师范学院学报．2000，23(5)：74-76
47 姜胜斌，廖桂英，袁华，王应席．液相催化氧化乳酸制丙酮酸．上海化工．2001，3，4：29-31
48 EisenbergA, Seip J E, Gavagan J E et al. Pyruvic acid production using methylotrophic yeast transformants as catalyst. J Molecular Catalysis B-Enzymatic. 1997,2:223-232
49 Shimizu A, Ogawa O. Manufacturing method for pyruvic acid . Japanese Patent, JP 11,243,979. 1999
50 Miyata R, Yonehara T, Yomoto K. Manufacture of pyruvic acid from L-tartaric acid with Pseudomonas putida. Japanese Patent, JP 61,216,695, 1986
51 Tarama S , Sogabe Y , Mitsuhida N . Manufacture of pyruvic acid with enzymes or bacteria. JP 63,133 ,990,1988
52 Anton D L, Dicosimo R, Witterholt V G. Process for the preparation of pyruvic acid. WO patent 9,500,656,1995
53 Anton D L, Dicosimo R, Witterholt V G. Production of pyruvic acid using pemeabilized transformants of Hansenula polymorpha and Pichia pastoris which express glycolate oxidase and catalase. US Patent ,No5,538,875,1996
54 GU J S (谷劲松),XU p(许平), LI T L(李铁林). Preparation of pyruvate from lactate oxidase. Chinese Joural of Applied and EnvironMental Biology(应用与环境生物学报). 2001,7 (6) : 616-620
55 Schinschel C, Simon H. Preparation of pyruvate from (R)-lactate with Proteus species. J Biotechnol.1993,31:191-203
56 Gunther H, Neumann S, Simon H. 2-oxocarboxylate reductase from Proteus species and its use for the preparation of (2R)-hydroxy acids, J Biotechnol. 1987, 5:53-65
57 Izumi Y, Matsumura Y, Tani Y et al. Pyruvic acid production from 1,2-propandediol by thiamine-requiring Acinetobacter sp 80-M. AgricBiol Chem. 1982 , 46:2673-2679
58 Moriguchi M, Shuto K, Hashimoto T. Production of pyruvic acid from saccharified citrus peel extract by dried ceels of Debaryomyces coudertii. J Ferment Technol.1984,62:243-248
59 Miyata R, Tsutsui H, Yonehara T. Manufacture of pyruvic acid with Torulopsis species. Janpaese Patent, JP O,155,185,1989
60 Yonehara T, Miyata R et al. Development of fermentative production of pyruvate by metabolic control-Monograph.Seibutsu Kogaku Kaishi (in Japanese) 2000,78:56-62
61 Yokota A, lota S, Takao S. Tryptophan production by a lipoic acid auxotroph Enterobacter aerogenes having both pyruvic.acid productivity and high tryptophanase activity. Sgric Biol chem. 1989,53:2037-2044
62 Yanse H, Mori N, Masuda M et al. Pyruvate production by Enterococcus casseliflavus A-12 from gluconate in an alkaline medium. J Ferment Bioeng, 1992,73:287-291
63 Yokota A, Terasawa Y, Takaota N et al. Pyruvate production by an F_1-ATPase-defective mutant of Escherichia Coli. Biosci Biotechnol Biochem.1994,58:2164-2167
64 Besnainou B, Giani D, Sahut C. Process for the production of pyruvic acid by fermentation. European Patent. No EP 312.453,1989.
65 Takao Stanida M. Pyruvic acid production by Schizophyllum commune. J Ferment Technol. 1982,60:277-280
66 Uchio R,Kikuchi K, Hirose Y. Process for producing pyruvic acid by fermentation. US Patent, No. 3,993,543,1976
67 Miyata R, Yonehara T. Improvement of fermentative production of pyruvate from glucose by Torulopsis glabrata IF00005. J Ferment Bioeng.1996,82:475-479
68 Miyata R. Yonehara T. Fermentative manufacture of pyruvic acid and its salts. Janpanese Patent. JP 09,047,292,1997
69 LIU L M(刘立明), LI Y(李寅), DU G C(堵国成) et al. Breeding of high-pyruvate-producting Torulopsis glabrata with acetate as supplement carbon source. Industrial Microbiology (工业微生物) . 2002, 32(3):10-14
70 沈同，王静岩．生物化学．第2版，高等教育出版社．1990年
71 李寅，陈坚，伦世仪．维生素在丙酮酸过量合成中的重要作用．微生物学报．2000，40(5)：528-534
72 刘立明，李寅，堵国成．乙酸渗漏型丙酮酸高产菌的选育．工业微生物．2002，32(3)：10-14
73 李寅，陈坚，陈燕．丙酮酸高产菌株的选育及中试研究．工业微生物．2001，31(2)：10-13
74 陈燕，李寅，陈坚．葡萄糖对光滑球拟酵母发酵生产丙酮酸的影响．无锡轻工大学学报．2001，20(2)：137-141
75 李寅，陈坚，梁大芳．营养条件对光滑球拟酵母发酵生产丙酮酸的影响．生物工程学报．2000，16(2)：225-228
76 袁辉，华子春．丙酮酸野生酵母菌的筛选及其生理生化特性研究．微生物学杂
志．2001，21(3)：12-14
77 姚忠，李家璜，欧阳平凯．丙酮酸化学萃取的盐效应．高校化学工程学报．2001，15(6)：587-590
78 李寅，傅为民，陈坚．离子交换法从发酵液中提取丙酮酸．无锡轻工大学学报．2001，20(4)：335-339
79 叶应妩，李建斋，王玉深．临床实验诊断学(上册)．北京：人民卫生出版社．1989，439-469
80 Lamperecht W, Heinz F. In:Methods of Enzymatic Analysis, ed. Bergmeyer, H.U. VCH. Weinheim. 1984,6:570-577
81 朱永和，何金柱．血液中乳酸、丙酮酸琥珀酸的气相色谱微量分析法．中华医学检验杂志．1987，10(5)：275-276
82 倪星忠．临床生化酶试剂方法．上海：华东师范大学出版社．1993，127-128
83 梁倩玲，刘丽芳．HPLC法测定丙酮酸钙的含量．广州化学．2002，27(1)：29-31
84 蒋淑艳，曲复宁．同步荧光—双波长法同时测定抗坏血酸和丙酮酸．分析化学．25(9)：1064-1067
85 潘振球，陈波，王春娥，冯家力，伍碧珍．离子排阻色谱—紫外检测分析复配方保健食品中丙酮酸钙含量．中国公共卫生．2001，17(6)：576
86 谭宁宁，王荣珍，曹维娟．呼吸道感染患儿血液丙酮酸含量测定及临床观察．江苏医药．1996，22(2)：116
87 刘振宝．精液丙酮酸含量分析．蚌埠医学院学报．1994，19(3)：260-261
88 Miller. G.L. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem.. 1959,31(3): 426-428
2 段长强，孟庆芳，张泰等．现代化学试剂手册(第一分册)[M]．北京：化学工业出版社，1988
3 潘竹林．丙酮酸缺陷症研究现状．中华血液学杂志．1999，20(4)：223-224
4 唐胜南，李艳，伍期专．血乳糖、丙酮酸比值测定的意义．中国卫生检验杂志．1999，9(6)：455-456
5 谭宁宁，王荣珍，曹维娟．呼吸道感染患儿血液丙酮酸含量测定及临床观察．江苏医药．1996，22(2)：116
6 Yonehara T. Miyata R. Fermentative production of pyruvate from glucose by Torulgpsis glabrata. J Ferment Bioeng. 1994,78:155-159
7 Nagasawa T, Utagawa T, Goto J, Kim CJ, Tani Y, Kumagai H, Yamada H. Synthesis of L-tyrosine-related amino acids by tyrosine phenol-lyase of Citrobacter intermedius Eur J Biochem.1981, 117:33-40
8 姜胜斌，廖桂英，袁华．丙酮酸系列产品．精细与专用化学品．2000，21：8-9
9 袁辉，华子春．丙酮酸发酵能力的提高．微生物学杂志．2000，20(3)：23-24，39
10 Stanko, R.T., Robertson, R.J., Spina, R.J., ReAlly, J.J., Greenawalt, K.D., Goss, F.L. Enhancement of arm-exercise endurance capacity with dihydroxyacetone and pyruvate. Journal of Applied Physiology. 1990,68:119-124
11 Stanko, R.T., Robertson, R.J., Galbreath, R.W., ReAlly, J.J., Greenawalt, K.D., Goss, F.L. Enhanced leg-exercise endurance with a high-carbohydrate diet and dihydroxyacetone and pyruvate. Journal of Applied Physiology. 1990,69:1651-1656
12 Robertson, KJ., Stanko, R.T., ReAlly, J.J., Greenawalt, K.D., Goss, F.L., Spina, R.J. Blood glucose extraction as a mediator of perceived exertion during prolonged exercise. European Journal of Applied Physiology. 1990,61:100-105
13 Stanko, RT. Tietze, D.L., Arch, J.E. Body composition, energy utilization, and nitrogen metabolism with a 4.25-MJ/d low-energy diet supplemented with pyruvate. American Journal of Clinical Nutrition. 1992,56:630-635
14 Stanko, R.T. Tietze, D.L., Arch, i.E. Body composition, energy utilization and nitrogen metabolism with a severely restricted diet supplemented with dihydroxyacetone and pyruvate. American Journal of Clinical Nutrition. 1992,55:771-775
15 Stanko, R.T., Adibi, S.A. Inhibition of lipid accumulation and enhancement of energy expenditure by the addition of pyruvate and dihydroxyacetone to a rat diet. Metabolism. 1986,35:182-186
16 Stanko, R.T., Ferguson, T.L., Newman, C.W., Newman, R.K. Reduction of carcass fat in swine with dietary addition of dihydroxyacetone and pyruvate. Journal of Animal Science. 1989,6:1272-1278
17 Cortez, M.Y., Torgan, C.E., Brozinick, J.T., Miller, R.H., Ivy, J.L. Effects of pyruvate and dihydroxyacetone consumption on the growth and metabolic state of obese Zucker rats. American Journal of Clinical Nutrition. 1991,53:847-853
18 James B. Roufs, M.S., Pyruvate—Does It Amp Endurance and Burn More Fat?", Muscle & Fitness Magazine. Dec. 1996
19 Andy Calhoun, Pyruvate: The Universal Supplement, The Athlete's Power Source Quarterly Winter 97-98
20 吕咏梅．丙酮酸钙及原料丙酮酸发展前景广阔．中国医药情报．2002，8(3)：52-53
21 张庆勇．丙酮酸及丙酮酸钠的制备．化学试剂．1997，19(6)：375，336
22 精细化学产品目录[M]．化学试剂．北京：化学工业出版社．2000
23 傅月华，余炳辉，纪佩芳．应用丙酮酸钠检出食品中亚致死菌的研究．12-14
24 Howard J W, Fraser W A. Preparation of pyruvic acid. Org Synth Coll.1932,1:475-480
25 US 4 136 111,1979
26 JP 7 981 221,1979
27 JP 9 085 230,1990
28 JP 7 939 016,1979
29 吴俊明．1，2-丙二醇空气氧化合成丙酮醛[J]．化工时刊．1997，11(12)：26-28
30 朱京．丙二醇空气氧化制丙酮醛的电结晶银催化剂[J]．复且大学学报．1989，28(1)：14-18
31 Get 3 219 355, 1983
32 Baizer M M. [J]. Tetrahedron. 1984, 40(6): 935-969
33 Chaussard J, Folest J C, et al [J]. Synthesis. 1990,5:369-381
34 廖桂英，姜肚斌，袁华．丙酮酸的电化学合成法．天津化工．2000，6：29
35 UK 2 018 773. 1979
36 J Mol. Catal [J].1992, 71(1):25-35
37 JP 5 619 854,1981
38 JP 5 724 336, 1982
39 Mamoru Ai,.Kyoji Ohdan. Chem Lett[J].1995, (5): 405
40 Mamoru Ai, Kyoji Ohdan. APPL. Catal A. 1997,150(1): 13-20
41 USP 4 229 590, 1980
42 JP 8 697 247,1986
43 胡旭灿等．乳酸乙脂在Ag／NaY催化剂上的脱氢反应制丙酮酸乙酯[J]．催化学报．1998，19(5)：428-431
44 杨辉琼，易翔．乳酸氧气氧化法制备丙酮酸．化学世界．2002，43(6)：307-309
45 杨辉琼，郭贤烙，易翔．双氧水催化氧化乳酸合成丙酮酸．化学工程师．2002，2：14-15
46 廖桂英，王应席．以磷酸铁盐为催化剂：乳酸氧化脱氢制丙酮酸．荆州师范学院学报．2000，23(5)：74-76
47 姜胜斌，廖桂英，袁华，王应席．液相催化氧化乳酸制丙酮酸．上海化工．2001，3，4：29-31
48 EisenbergA, Seip J E, Gavagan J E et al. Pyruvic acid production using methylotrophic yeast transformants as catalyst. J Molecular Catalysis B-Enzymatic. 1997,2:223-232
49 Shimizu A, Ogawa O. Manufacturing method for pyruvic acid . Japanese Patent, JP 11,243,979. 1999
50 Miyata R, Yonehara T, Yomoto K. Manufacture of pyruvic acid from L-tartaric acid with Pseudomonas putida. Japanese Patent, JP 61,216,695, 1986
51 Tarama S , Sogabe Y , Mitsuhida N . Manufacture of pyruvic acid with enzymes or bacteria. JP 63,133 ,990,1988
52 Anton D L, Dicosimo R, Witterholt V G. Process for the preparation of pyruvic acid. WO patent 9,500,656,1995
53 Anton D L, Dicosimo R, Witterholt V G. Production of pyruvic acid using pemeabilized transformants of Hansenula polymorpha and Pichia pastoris which express glycolate oxidase and catalase. US Patent ,No5,538,875,1996
54 GU J S (谷劲松),XU p(许平), LI T L(李铁林). Preparation of pyruvate from lactate oxidase. Chinese Joural of Applied and EnvironMental Biology(应用与环境生物学报). 2001,7 (6) : 616-620
55 Schinschel C, Simon H. Preparation of pyruvate from (R)-lactate with Proteus species. J Biotechnol.1993,31:191-203
56 Gunther H, Neumann S, Simon H. 2-oxocarboxylate reductase from Proteus species and its use for the preparation of (2R)-hydroxy acids, J Biotechnol. 1987, 5:53-65
57 Izumi Y, Matsumura Y, Tani Y et al. Pyruvic acid production from 1,2-propandediol by thiamine-requiring Acinetobacter sp 80-M. AgricBiol Chem. 1982 , 46:2673-2679
58 Moriguchi M, Shuto K, Hashimoto T. Production of pyruvic acid from saccharified citrus peel extract by dried ceels of Debaryomyces coudertii. J Ferment Technol.1984,62:243-248
59 Miyata R, Tsutsui H, Yonehara T. Manufacture of pyruvic acid with Torulopsis species. Janpaese Patent, JP O,155,185,1989
60 Yonehara T, Miyata R et al. Development of fermentative production of pyruvate by metabolic control-Monograph.Seibutsu Kogaku Kaishi (in Japanese) 2000,78:56-62
61 Yokota A, lota S, Takao S. Tryptophan production by a lipoic acid auxotroph Enterobacter aerogenes having both pyruvic.acid productivity and high tryptophanase activity. Sgric Biol chem. 1989,53:2037-2044
62 Yanse H, Mori N, Masuda M et al. Pyruvate production by Enterococcus casseliflavus A-12 from gluconate in an alkaline medium. J Ferment Bioeng, 1992,73:287-291
63 Yokota A, Terasawa Y, Takaota N et al. Pyruvate production by an F_1-ATPase-defective mutant of Escherichia Coli. Biosci Biotechnol Biochem.1994,58:2164-2167
64 Besnainou B, Giani D, Sahut C. Process for the production of pyruvic acid by fermentation. European Patent. No EP 312.453,1989.
65 Takao Stanida M. Pyruvic acid production by Schizophyllum commune. J Ferment Technol. 1982,60:277-280
66 Uchio R,Kikuchi K, Hirose Y. Process for producing pyruvic acid by fermentation. US Patent, No. 3,993,543,1976
67 Miyata R, Yonehara T. Improvement of fermentative production of pyruvate from glucose by Torulopsis glabrata IF00005. J Ferment Bioeng.1996,82:475-479
68 Miyata R. Yonehara T. Fermentative manufacture of pyruvic acid and its salts. Janpanese Patent. JP 09,047,292,1997
69 LIU L M(刘立明), LI Y(李寅), DU G C(堵国成) et al. Breeding of high-pyruvate-producting Torulopsis glabrata with acetate as supplement carbon source. Industrial Microbiology (工业微生物) . 2002, 32(3):10-14
70 沈同，王静岩．生物化学．第2版，高等教育出版社．1990年
71 李寅，陈坚，伦世仪．维生素在丙酮酸过量合成中的重要作用．微生物学报．2000，40(5)：528-534
72 刘立明，李寅，堵国成．乙酸渗漏型丙酮酸高产菌的选育．工业微生物．2002，32(3)：10-14
73 李寅，陈坚，陈燕．丙酮酸高产菌株的选育及中试研究．工业微生物．2001，31(2)：10-13
74 陈燕，李寅，陈坚．葡萄糖对光滑球拟酵母发酵生产丙酮酸的影响．无锡轻工大学学报．2001，20(2)：137-141
75 李寅，陈坚，梁大芳．营养条件对光滑球拟酵母发酵生产丙酮酸的影响．生物工程学报．2000，16(2)：225-228
76 袁辉，华子春．丙酮酸野生酵母菌的筛选及其生理生化特性研究．微生物学杂
志．2001，21(3)：12-14
77 姚忠，李家璜，欧阳平凯．丙酮酸化学萃取的盐效应．高校化学工程学报．2001，15(6)：587-590
78 李寅，傅为民，陈坚．离子交换法从发酵液中提取丙酮酸．无锡轻工大学学报．2001，20(4)：335-339
79 叶应妩，李建斋，王玉深．临床实验诊断学(上册)．北京：人民卫生出版社．1989，439-469
80 Lamperecht W, Heinz F. In:Methods of Enzymatic Analysis, ed. Bergmeyer, H.U. VCH. Weinheim. 1984,6:570-577
81 朱永和，何金柱．血液中乳酸、丙酮酸琥珀酸的气相色谱微量分析法．中华医学检验杂志．1987，10(5)：275-276
82 倪星忠．临床生化酶试剂方法．上海：华东师范大学出版社．1993，127-128
83 梁倩玲，刘丽芳．HPLC法测定丙酮酸钙的含量．广州化学．2002，27(1)：29-31
84 蒋淑艳，曲复宁．同步荧光—双波长法同时测定抗坏血酸和丙酮酸．分析化学．25(9)：1064-1067
85 潘振球，陈波，王春娥，冯家力，伍碧珍．离子排阻色谱—紫外检测分析复配方保健食品中丙酮酸钙含量．中国公共卫生．2001，17(6)：576
86 谭宁宁，王荣珍，曹维娟．呼吸道感染患儿血液丙酮酸含量测定及临床观察．江苏医药．1996，22(2)：116
87 刘振宝．精液丙酮酸含量分析．蚌埠医学院学报．1994，19(3)：260-261
88 Miller. G.L. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem.. 1959,31(3): 426-428