藻蓝蛋白及其水解物促进玉米直支链淀粉回生机理研究
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摘要
藻蓝蛋白是螺旋藻中对人体具有多种保健功能的蛋白,初步发现其具有明显促进玉米淀粉回生的作用。为了进一步探索其促进玉米淀粉回生的机理,该文将1%和10%藻蓝蛋白及其水解物添加至玉米直链、支链及其混合淀粉中,测定其对淀粉回生的影响,通过X-射线衍射、差示扫描量热、红外和固体核磁分析藻蓝蛋白及其水解物促进玉米淀粉回生机理。研究结果表明,添加1.0%藻蓝蛋白对直链淀粉回生率没有影响,使支链淀粉回生率提高了61.4%;而添加量为10%的藻蓝蛋白使直链淀粉回生率提高了60.4%,使支链淀粉回生率提高了69.6%。藻蓝蛋白水解肽低的添加量(1.0%)对玉米直链的回生率影响不显著,但使支链淀粉的回生提高了28.1%;水解肽高添加量(10.0%)使玉米直链淀粉的回生率提高了184.7%,使玉米支链淀粉的回生率提高了47.0%。紫外可见扫描显示碱性蛋白酶水解可使处于藻蓝蛋白结构中心的藻蓝素外露出来。X射线结果表明藻蓝蛋白与玉米直链支链淀粉混合回生后分别产生一个衍射角为2θ为16.44o、16.60o尖锐衍射峰。差热结果显示,藻蓝蛋白与玉米支链淀粉混合、藻蓝蛋白水解物与玉米直链淀粉混合后,所得回生淀粉均失去结晶峰,而出现唯一重结晶峰。红外和~(13)C固体核磁结果表明,藻蓝蛋白通过精氨酸氨基与玉米支链淀粉还原端醛基形成氢键,利用藻蓝蛋白中疏水性氨基酸驱离玉米支链淀粉侧链末端水分子,促使支链淀粉末端双螺旋解旋,加快了支链淀粉末端链间氢键形成,提高了其回生率;而藻蓝蛋白水解物中半胱氨酸巯基和玉米直链淀粉还原端醛基在与直链淀粉混合回生过程形成氢键,通过分子甩动使直链淀粉双螺旋解开,大大促进了玉米直链淀粉间形成氢键,提高了其回生率。该研究提供了一种提高玉米淀粉回生率的新技术。
Phycocyanin is a kind of dark blue powder isolated from spirulina and has a variety of health functions of anti-cancer, blood cell regeneration, etc. In order to widen its application in food, phycocyanin and its hydrolysis are added into maize amylose/amylopectin paste during retrogradation, it has been found that they can promote the retrogradation of amylose/amylopectin. In order to further explore its mechanism of promoting the retrogradation of maize starch, the effects of contents of phycocyanin or its hydrolysate in maize amylose/amylopectin on retrogradation rate were determined, and the method of X-ray diffraction, differential scanning calorimetry, infrared and solid nuclear magnetic analysis were used. The results showed that phycocyanin hydrolyzate promoted maize amylose retrogradation more obviously than that of phycocyanin. 10% addition of phycocyanin hydrolyzate could make maize amylose retrogradation rate increase from 27% to 76.9%, 184.8% higher. Phycocyanin promoted the retrogradation of maize amylopectin more pronouned than its hydrolysate, 10% addition of phycocyanin let maize amylopectin retrogradation rate increase from 26.7% to 45.3%, which increased by 69.6%. According to the UV-visible scanning, phycocyanin hydrolysis co cause the phycocyanin pigment in the center of phycocyanin to be exposed. X-ray results indicated that addition of phycocyanin to maize amylopectin induced a sharper peak at 2θ is 16.60o, as well as that mixture of phycocyanin hydrolysate and maize amylose caused a same sharper one at 2θ is 16.44o. Compared to diffraction angles of retrograded maize amylose/amylopectin, the decrease of sharp diffraction angle in those mixtures supported that interplanar distance at this angle became larger. DSC results demonstrated that mixture of phycocyanin and maize amylopectin, or mixture of phycocyanin hydrolysate and maize amylose, which both promoted retrogradation greatly, both caused the presence of re-crystallization peaks and loss of crystallization peaks. In other words, more phycocyanin/hydrolysate chains and maize amylose/amylopectin with similar length involved in retrogradation. Such process should deal with unwinding of maize amylose/amylopectin. The results of infrared and 13 C solid NMR showed that phycocyanin might be hydrolyzed into arginine-rich polypeptide with hydrophilic, polysaccharides and polypeptide with hydrophobic. The mechanism for phycocyanin to promote retrogradation of maize amylopectin was that the formation of hydrogen bond between the arginine amino acid of phycocyanin and aldehyde of reduction end in maize amylopectin, and water molecule of maize amylopectin being driving away from the side chain end by phycocyanin hydrophobic amino acid, promoted the end of the chain-branched starch double helix spin, accelerate the formation of hydrogen bond between the end chains of amylopectin, which lead to improve retrogradation rate of maize amylopectin. The mechanism for phycocyanin hydrolysate to increase retrogradation rate of maize amylose was that the hydrogen bond formed between the sulfydryl of cysteine in phycocyanin hydrolyzate and aldehyde of reduction end in maize amylose during retrogradation of mixture, then the double helix of amylose was unlocked by molecular swinging, which greatly promoted the formation of hydrogen bond between amylose and improved the retrogradation rate of amylose. This study provides a new technique to improve the retrogradation rate of maize starch.
Phycocyanin is a kind of dark blue powder isolated from spirulina and has a variety of health functions of anti-cancer, blood cell regeneration, etc. In order to widen its application in food, phycocyanin and its hydrolysis are added into maize amylose/amylopectin paste during retrogradation, it has been found that they can promote the retrogradation of amylose/amylopectin. In order to further explore its mechanism of promoting the retrogradation of maize starch, the effects of contents of phycocyanin or its hydrolysate in maize amylose/amylopectin on retrogradation rate were determined, and the method of X-ray diffraction, differential scanning calorimetry, infrared and solid nuclear magnetic analysis were used. The results showed that phycocyanin hydrolyzate promoted maize amylose retrogradation more obviously than that of phycocyanin. 10% addition of phycocyanin hydrolyzate could make maize amylose retrogradation rate increase from 27% to 76.9%, 184.8% higher. Phycocyanin promoted the retrogradation of maize amylopectin more pronouned than its hydrolysate, 10% addition of phycocyanin let maize amylopectin retrogradation rate increase from 26.7% to 45.3%, which increased by 69.6%. According to the UV-visible scanning, phycocyanin hydrolysis co cause the phycocyanin pigment in the center of phycocyanin to be exposed. X-ray results indicated that addition of phycocyanin to maize amylopectin induced a sharper peak at 2θ is 16.60o, as well as that mixture of phycocyanin hydrolysate and maize amylose caused a same sharper one at 2θ is 16.44o. Compared to diffraction angles of retrograded maize amylose/amylopectin, the decrease of sharp diffraction angle in those mixtures supported that interplanar distance at this angle became larger. DSC results demonstrated that mixture of phycocyanin and maize amylopectin, or mixture of phycocyanin hydrolysate and maize amylose, which both promoted retrogradation greatly, both caused the presence of re-crystallization peaks and loss of crystallization peaks. In other words, more phycocyanin/hydrolysate chains and maize amylose/amylopectin with similar length involved in retrogradation. Such process should deal with unwinding of maize amylose/amylopectin. The results of infrared and 13 C solid NMR showed that phycocyanin might be hydrolyzed into arginine-rich polypeptide with hydrophilic, polysaccharides and polypeptide with hydrophobic. The mechanism for phycocyanin to promote retrogradation of maize amylopectin was that the formation of hydrogen bond between the arginine amino acid of phycocyanin and aldehyde of reduction end in maize amylopectin, and water molecule of maize amylopectin being driving away from the side chain end by phycocyanin hydrophobic amino acid, promoted the end of the chain-branched starch double helix spin, accelerate the formation of hydrogen bond between the end chains of amylopectin, which lead to improve retrogradation rate of maize amylopectin. The mechanism for phycocyanin hydrolysate to increase retrogradation rate of maize amylose was that the hydrogen bond formed between the sulfydryl of cysteine in phycocyanin hydrolyzate and aldehyde of reduction end in maize amylose during retrogradation of mixture, then the double helix of amylose was unlocked by molecular swinging, which greatly promoted the formation of hydrogen bond between amylose and improved the retrogradation rate of amylose. This study provides a new technique to improve the retrogradation rate of maize starch.
引文
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[2]张坤生,周雪,连喜军,等.冷藏条件下荞麦淀粉回生规律的研究[J].食品工业科技,2013,34(16):157-158,177.Zhang Kunsheng,Zhou Xue,Lian Xijun,et al.The study on the retrogadation regular of buckwheat starch under cold storage condition[J].Science and Technology of Food Industry(STFI),2013,34(16):157-158,177.(in Chinese with English abstract)
[3]张坤生,宁仲娟,连喜军,等.冷冻对糯米淀粉回生的影响[J].食品工业科技,2013,34(21):49-51.Zhang Kunsheng,Ning Zhongjuan,Lian Xijun,et al.Effect of freezing on starch retrogradation of glutinous rice starch[J].Science and Technology of Food Industry(STFI),2013,34(21):49-51.(in Chinese with English abstract)
[4]连喜军,张燕.不同老化工艺对甘薯回生抗性淀粉制备率的影响[J].粮食加工,2012,37(3):46-48.Lian Xijun,Zhang Yan.Effects of different aging processes on the preparation rate of resistant starch of sweet potato[J].Grain Processing,2012,37(3):46-48.(in Chinese with English abstract)
[5]Masatcioglu T M,Sumer Z,Koksel H.An innovative approach for significantly increasing enzyme resistant starch type 3 content in high amylose starches by using extrusion cooking[J].Journal of Cereal Science,2017,74:95-102.
[6]Jagannadham K,Parimalavalli R,Babu A S.Effect of triple retrogradation treatment on chickpea resistant starch formation and its characterization[J].Journal of Food Science and Technology,2017,54(4):901-908.
[7]Silverio J,Fredriksson H,Andersson R,et al.The effect of temperature cycling on the amylopectin retrogradation of starches with different amylopectin unit-chain length distribution[J].Carbohydrate Polymers,2000,42(2):175-184.
[8]连喜军,罗庆丰,刘学燕,等.超声波对甘薯回生抗性淀粉生成的作用[J].食品研究与开发,2011,32(1):61-64.Lian Xijun,Luo Qingfeng,Liu Xueyan,et al.Effects of ultrasonic wave on the formation of resistant starch of sweet potato[J].Food Research and Development,2011,32(1):61-64.(in Chinese with English abstract)
[9]连喜军,钱瑞,刘敬,等.电解法和微波法联合处理提高甘薯淀粉回生率[J].粮食加工,2011,36(5):51-54.Lian Xijun,Qian Rui,Liu Jing,et al.The retrogradation rate of sweet potato starch was improved by electrolysis and microwave[J].Food Research and Development,2011,36(5):51-54.(in Chinese with English abstract)
[10]Demirkesen-Bicak H,Tacer-Caba Z,Nilufer-Erdil D.Pullulanase treatments to increase resistant starch content of black chickpea(Cicer arietinum L.)starch and the effects on starch properties[J].International journal of Biological Macromolecules,2018,111:505-513.
[11]连喜军,赵璐,牛瑞华,等.中温?-淀粉酶处理提高甘薯回生抗性淀粉制备率[J].粮食与油脂,2010(8):21-23.Lian Xijun,Zhao Lu,Niu Ruihua,et al.The preparation rate of retrograde resistant starch of sweet potato was improved by medium temperature and?-amylase treatment[J].Cereals&Oils,2010(8):21-23.(in Chinese with English abstract)
[12]丁文平,李清,夏文水.淀粉酶对大米淀粉回生影响机理的研究[J].粮食与饲料工业,2005(10):16-17.Ding Wenping,Li Qing,Xia Wenshui.Study on the mechanism of amylase on the retrogradation of rice starch[J].Cereal&Feed Industry,2005(10):16-17.(in Chinese with English abstract)
[13]孟宪昉,刘立增,郭俊杰,等.草酸侵蚀马铃薯回生淀粉制备晶种促进玉米淀粉回生的研究[J].食品工业科技,2016,37(3):131-134,148.Meng Xianfang,Liu Lizeng,Guo Junjie,et al.Study on the effect of oxalic acid on the retrogradation of corn starch by seed preparation of potato starch[J].Science and Technology of Food Industry(STFI),2016,37(3):131-134,148.(in Chinese with English abstract)
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