Fund-Nutritional Properties of Physiological Functions of Vitamin Biz and Antioxidative Substances of Algae

维生素Biz生理功能和藻类抗氧化物质的营养特性

• 批准号: 10680168
• 负责人: TAMURA Yoshiyuki
• 金额: $ 2.05万
• 依托单位: Hagoromo Gakuen Junior College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10680168/
• 关键词: chemiluminescence; vitamin B_<12> analogue; antioxidant; Porphyra yezoensis; vitamin B_<12>-deficientrat; methylmakonic acid; anserine; usujilene; ビタミンB_<12>; 補酵素型B_<12>; 過酸化ペルオキシド

Vitamin B_<12> concentrations of dried green (Enteromorpha sp.) and purple (Porphyra sp.) Iavers (nori) were determined by both Lactobacillus leichmannii ATCC 7830 microbiological and intrinsic factor chemiluminescence methods. The values determined by using the microbiological method (68.58±2.9 and 32.26±1.61 u g/1OOg of dry weight) were identical to (bund by using the chemiluminescence method (60.20±2.21 and 25.07±0.54 <μg/100g dry weight) in both dried green and purple lavers, respectively. The vitamin B_<12> concentration of an algal health food, spirulina (Spirulina sp.) tablets, was determined. The values determined with the microbiological method were 〜6-9-fold greater in the spirulina tablets than the values determined with chemiluminescence method. The contents of vitamin B_<12> in foods were assayed by using a fully automated chemiluminescence analyzer, which was compared with a microbiological method. In shellfishes and spirulina, the values determined by the microbiological … More method were〜6-8 told greater than the values determined by the chemiluminescence method, although there was good similarity between the values by the two methods in other foods. To clarify the bioavailability of vitamin B_<12> in lyophilized purple laver(nori), total vitamin B_<12> and vitamin B_<12> analogue contents were determined, and the effects of feeding the laver to vitamin B_<12>-deficient rats were investigated. The purple laver contained five types of biologically active vitamin B_<12> compounds(cyano-, hydroxo-, sulfito-, adenosyl-, and methylcobalamin), in which vitamin B_<12> coenzymes (adenosyl- and rnethylcobalamin) comprised about 60% of the total vitamin B_<12>. When 9-week-old vitamin Burdeficient rats, which excreted substantial amounts of methylmaIonic acid(71.7(SE 20.2) μ mol/d) in urine, were fed the diet purple laver (10μg/kg diet) for 20 d. urinary methylmalonic acid excretion (as an index of vitamin B_<12> deficiency) become uncletectable and hepatic vitamin B_<12> (especially adenosylcobalamin) levels were significantly increased. These results indicate that vitamin B_<12> in dried purple laver is bioavailable to rats. Dunaliella tertiolecta neither required vitamin B_<12> for growth nor took up exogenous vitamin B_<12> significantly. The Dunaliella cells grown in a vitamin B_<12>-free medium contained biologically active vitamin B_<12> (20.18±5.53 ng/g wet weight). About 77% of vitamin B_<12> found in the cells was recovered in macromolecular fractions (adenosyl- B_<12>, 39% ; and methyl- B_<12>, 21%). These results suggest that the alga has ability to synthesize vitamin B_<12> coenxymes.To examine the antioxidative compounds of nonpmtein amino acids in a red alga. Porphyra yezoensis, an ethanol extract of the cultured thalli was fractionated with Dowex column. The basic fraction V showed strong antioxidative capacities with ferric thiocyanate and TBARS measurements. In this basic fraction, histidine, 3-methylhistidine, carnosine, and anserine wewre detected beside ornitine, lysine, and arginine by amino acid analysis. The occurrence ofcarnosine and anserine suggests that the histidine-related compounds also contribute to the antioxidative reactions in P. yezoensis. The antioxidant activities of several extracts from Susabinori were measured by the ferric thiocyanate method and the thiobarbituric acid method. The methanol, acetone, ethyl acetate, and Ivexane extracts, and the chloroform-soluble and water-soluble fractions from the chloroform-metanol extract exhibited higher activities than a -tocopherol. The hot water extract showed little activity. Thin-layer chromatography analysis of the active extracts suggested the existence of several antioxidants. The activity of the chloroform soluble fraction was due to chlorophyll analogs. A strong antioxiclant was isolated from the methanol extract, accompanied by several amino acids such as leucine and phenylalanine. This compound was identified as usujilene, a kind ofmycosporine-glycine like amino acid. Less

维生素B_<12>干绿色（浒苔属）的浓度和紫色（Porphyra sp.）通过莱氏乳杆菌ATCC 7830微生物化学发光法和内因子化学发光法测定紫菜。用微生物法测定干绿色紫菜和干紫紫菜的含量（68.58±2.9和32.26±1.61 μ g/100 g干重）与化学发光法测定的含量（60.20±2.21和25.07±0.54 μg/100 g干重）基本一致。研究<12>了海藻保健食品螺旋藻（Spirulinasp.）片剂，已确定。螺旋藻片中微生物法测定值比化学发光法高1.6 ~ 9倍。采用<12>全自动化学发光分析仪测定食品中维生素B_的含量，并与微生物法进行比较。在贝类和螺旋藻中，由微生物测定的值 ...更多信息 方法测定的值比化学发光法测定的值高出1.6 -8倍，尽管在其他食品中两种方法测定的值之间有很好的相似性。为明确紫菜冻干粉中维生素B_的生物利用度<12>，测定了紫菜冻干粉中总维生素B_<12>和维生素B_<12>类似物的含量，并研究了紫菜对维生素B<12>_缺乏大鼠的影响。紫紫菜中含有5种具有生物活性的维生素B_<12>化合物（氰基、羟基、硫代、腺苷和甲基钴胺素），其中维生素B_<12>辅酶（腺苷和甲基钴胺素）约占总维生素B_的60%<12>。给9周龄维生素B2缺乏的大鼠喂饲紫紫菜（10μg/kg饲料）20天，尿中甲基丙二酸的排泄量为71.7（SE 20.2）μ mol/d。尿中甲基丙二酸排泄（作为维生素B_缺乏的指标<12>）变得不可检测，肝脏维生素B_<12>（尤其是腺苷钴胺素）水平显著增加。说明干紫菜中维生素B_<12>对大鼠具有生物利用度。杜氏藻生长不需要维生素B_2<12>，也不显著吸收外源维生素B_<12>2。在不含维生素B_的培养基中生长的杜氏藻细胞<12>含有生物活性维生素B_<12>（20.18±5.53 ng/g湿重）。在细胞中发现的维生素B_约有77%<12>以大分子级分回收（腺苷- B_<12>，39% ;和甲基- B_<12>，21%）。这些结果表明，红球藻具有合成维生素B_辅酶的能力<12>。条斑紫菜（Porphyra yezoensis）是条斑紫菜（Porphyra yezoensis）培养菌体的乙醇提取物。碱性组分V具有较强的抗氧化能力与硫氰酸铁和TBARS测量。通过氨基酸分析，在该碱性组分中，除了鸟氨酸、赖氨酸和精氨酸之外，还检测到组氨酸、3-甲基组氨酸、肌肽和鹅肌肽。肌肽和鹅肌肽的出现表明，组氨酸相关化合物也有助于条斑紫菜的抗氧化反应。采用硫氰酸铁法和硫代巴比妥酸法测定了几种Susabinori提取物的抗氧化活性。甲醇、丙酮、乙酸乙酯和Ivexane提取物以及来自氯仿-间醇提取物的氯仿可溶性和水溶性级分表现出比α-生育酚更高的活性。热水提取物几乎没有活性。活性提取物的薄层色谱分析表明存在几种抗氧化剂。氯仿可溶部分的活性是由于叶绿素类似物。从甲醇提取物中分离出一种强抗氧化剂，并含有几种氨基酸，如亮氨酸和苯丙氨酸。该化合物被鉴定为一种类似于真菌孢菌素-甘氨酸的氨基酸usujilene。少

项目成果

Shigeo TAKENAKA, Sumi SUGIYAMA, Shuhei EBARA, Emi, MIYAMOTO, Katsuo ABE, Yoshiyuki, TAMURA, Fumio WATANABE, Shingo TSUYAME, and Yoshihisa NAKANO: "Feeding dried purple laver (n0ri) to vitamin B_<12> Deficient rats significantly improves vitamin B_<12>Stat

Shigeo TAKENAKA、Sumi SUGIYAMA、Shuhei EBARA、Emi、MIYAMOTO、Katsuo ABE、Yoshiyuki、TAMURA、Fumio WATANABE、Shingo TSUYAME 和 Yoshihisa NAKANO：“给维生素 B_<12> 缺乏的大鼠喂食干紫菜（n0ri）可显着改善维生素 B_

Fumio WATANABE, Shigeo TAKANAKA, Hiromi KATSURA, Emi MIYAMOTO, Katsuo ABE, Yoshiyuki TAMURA, Toshiyuki NAKATSUKA, Yoshihisa NAKANO: "Characterization of a Vitamin B_<12> Compound in the Edible Purple Laver, (Parhyra yezoensis)"Bioscience, Biotechnology, a

Fumio WATANABE、Shigeo TAKANAKA、Hiromi KATSURA、Emi MIYAMOTO、Katsuo ABE、Yoshiyuki TAMURA、Toshiyuki NAKATSUKA、Yoshihisa NAKANO：“食用紫菜（Parhyra yezoensis）中维生素 B_<12> 化合物的表征”生物科学、生物技术、

Fumio WATANABE, Shigeo TAKENAKA, Hiromi KATSURA, S.A.M.Zakir Hussain MASUMDER, Katsuo ABE, Yoshiyuki NAKATSUKA: "Pseudovitamin B_<12> is the Pseudominant Cobmide of an Algal Health Food, Spirulina Tablets"Jpurnal of Agricultural and Food chemistry. 47(11)

Fumio WATANABE、Shigeo TAKENAKA、Hiromi KATSURA、S.A.M.Zakir Hussain MASUMDER、Katsuo ABE、Yoshiyuki NAKATSUKA：“伪维生素 B_<12> 是藻类保健食品螺旋藻片的伪显性化合物”《农业与食品化学杂志》。

Watanabe F.,Takenaka S.,Abe K.,Tamura Y.,& Nakano Y.: "Comparison of a Microbiological Assay and a Fully Automated Chemiluminescent System for the Determination of Vitamin B12 in Food" Journal of Agricultural and Food Chemistry. 46(4). 1433-1436 (1998)

渡边 F.、竹中 S.、阿部 K.、田村 Y.、

Yoshiyuki TAMURA, Shigeo TAKENAKA, Sumi SUGIYAMA, Reiko NAKAYAMA: "Occurrence of Anserine as an Antioxidative Dipeptide in Red Alga, Porphyra yezoensis"Bioscience, Biotechnology, and Biochemistry. 63(3). 563-561 (1998)

Yoshiyuki TAMURA、Shigeo TAKENAKA、Sumi SUGIYAMA、Reiko NAKAYAMA：“Anserine 作为红藻、条斑紫菜中的抗氧化二肽的出现”生物科学、生物技术和生物化学。