壳寡糖的抗氧化性能及其对鲜切苹果褐变的影响
摘要
鲜切果蔬由于新鲜、食用方便等优点越来越受到广大消费者的青睐,但是与完整果蔬相比,鲜切果蔬品质更容易劣变。随着人们安全意识的提高,化学合成保鲜剂的使用越来越受到限制,所以寻找安全、无毒副作用的天然保鲜剂成为研究的热点。壳寡糖作为具有生物安全性的氨基寡糖,具有抗氧化、抑菌等作用,本文就壳寡糖的体外抗氧化效果、对鲜切苹果褐变的影响以及在体外对多酚氧化酶(PPO)抑制作用做了系统性研究,具体内容如下:
1.研究了壳寡糖对自由基的清除作用。已有相关报道证明壳寡糖对自由基具有清除作用,本研究以常用的抗氧化剂抗坏血酸和半胱氨酸为对照,对比评价壳寡糖的抗氧化性能。研究发现,壳寡糖对DPPH自由基、羟自由基、超氧阴离子都具有较好的清除作用。壳寡糖对DPPH自由基的IC50为26.06μg/ml,抗坏血酸和半胱氨酸分别是2.47μg/ml和2.43μg/ml。壳寡糖对羟自由基的IC50为0.27mg/ml,抗坏血酸和半胱氨酸分别是0.04mg/ml、0.07mg/ml。虽然抗坏血酸和半胱氨酸在低浓度条件下对超氧阴离子的清除作用优于壳寡糖,但是抗坏血酸最大清除率是50%,半胱氨酸的最大清除率是26.89%,壳寡糖的最大清除率达到了58%,说明壳寡糖对超氧阴离子的最终清除率比较高。上述结果表明壳寡糖虽然对三种自由基具有一定的清除能力,但其清除能力比抗坏血酸和半胱氨酸弱。
2.研究了壳寡糖对鲜切苹果褐变的影响。将厚度为2.5mm的鲜切苹果分别在蒸馏水、1%壳寡糖溶液、1%抗坏血酸溶液中浸泡5min,用吸水纸吸干后在10h内测定各个指标变化,以蒸馏水为阴性对照,抗坏血酸为阳性对照。研究发现,壳寡糖能够较好的抑制鲜切苹果的褐变:褐变指数在4h内明显比对照组低,与抗坏血酸组相当;多酚氧化酶活性明显比蒸馏水处理组和抗坏血酸组都要低。壳寡糖能显著提高鲜切苹果过氧化物酶(POD)含量,在10h内POD含量远远高于对照组和抗坏血酸组;壳寡糖还可以维持鲜切苹果的品质:壳寡糖处理的鲜切苹果失重率比其它两组稍低;壳寡糖对鲜切苹果的总糖含量影响不大,但可以降低可滴定酸度,这样就相应提高了糖酸比;壳寡糖还能抑制鲜切苹果表面的细菌含量,在10h内壳寡糖处理的鲜切苹果表面细菌总含量明显低于其它两组。
3.研究了壳寡糖对PPO的抑制作用。由上面的研究知道壳寡糖具有防止褐变的作用,且降低了PPO活性。本研究是在体外从反应动力学角度探讨壳寡糖对PPO的抑制作用。研究发现壳寡糖在体外对PPO有抑制作用,且随着浓度的增大抑制率增大,在5mg/ml时对PPO的抑制率为77%。以不同浓度的PPO为横坐标,以反应速率为纵坐标作图,发现加入不同浓度壳寡糖后所得到的动力学曲线经过同一点,且随着壳寡糖浓度的增大,斜率变小,说明壳寡糖是PPO的可逆性抑制剂。不同浓度壳寡糖存在条件下获得的双倒数动力曲线图得到的是一组平行线,说明壳寡糖是PPO的可逆性反竞争抑制剂。
壳寡糖可抑制鲜切苹果果肉褐变,提高贮藏期间果肉品质。尽管壳寡糖对自由基的清除作用低于抗坏血酸,但壳寡糖显著抑制了PPO的活性,且通过抑制动力学的研究发现其作用类型为反竞争性抑制。本文结果表明壳寡糖主要可作为PPO的反竞争性抑制剂抑制鲜切苹果褐变的发生。
Due to freshness and convenience, Fresh-cut fruits and vegetables are becoming more and more popular with people of all ages. But compared with the perfect ones, the quality of fresh-cut fruits and vegetables is easy to deteriorate. The research of finding safe and non-toxic natural preservation has becoming a hot topic because of people’s safety awareness. OCS, as a biosafe amino polysaccharides, also have the properties of antioxidant and antimicrobial. This study is about the antioxidant of OCS, the effect of browning of fresh-cut apples and the inbition of PPO.The details are as follow.
1. The radical scavenging activity of OCS. There have been some reports about the radical scavenging activity of OCS. This study set the ASA and cysteine as the control. The results showed that OCS has a good scavenging activity on DPPH radical, hydroxyl radical and superoxide anion. The IC50 of OCS on DPPH radical scavenging activity is 26.06ug/ml, well the ASA is 2.47ug/ml, the cysteine is 2.43ug/ml. The IC50 of OCS on hydroxyl radical scavenging is 0.27mg/ml, well the ASA is 0.04mg/ml, the cysteine is 0.07mg/ml. Although ASA and cysteine can scaveng superoxide anion at a low concentration, they did’t have a thorough scavenging ability. The maximum clearance rate of ASA is 50%, the cysteine is 26.89%. The OCS can reach 58%. In short, the OCS has a lower radical scavenging activity compared with ASA and cysteine.
2. The effect of OCS on the browning of fresh-cut apples. The fresh-cut apples which have a thickness of 2.5mm has been separately dipped in solution of distilled water, 1%ASA, 1%OCS for 5min, then, been dryed by absorbent paper. After this treatment, the fresh-cut apples were used to measure all indicators about the qulity of the apples within 10h. All the results made the ASA as positive control, and made the distilled water as negative control. This study has found that oligocatosan can inhibit the browning of fresh-cut apples. Compared with the apples treated by distilled water, the browning index of apples treated by OCS was lower within 4 hours. The OCS treatment had a lower PPO activity than the ASA and distilled water treatment. The content of POD is significantly improved within 10h compared with the other treatments. OCS can also maintain the quality of fresh-cut apples. The weight loss of OCS group is slightly lower than the other groups. The OCS has little effect on total sugar content of fresh-cut apples, but can reduce the titratable acidity, so the sugar to acid ratio has been increased. OCS can inhibit the growth of bacteria on the surface of fresh-cut apples. The content of bacteria on apples treated by OCS is significantly lower than the other groups.
3. The inhibition effect from OCS on PPO. From the study above, we know the OCS can inhibit browning and reduce the activity of PPO. The study about the inhibition from OCS on PPO is working through the method of reaction dynamics in vitro. The OCS can inhibit the activity of PPO, and the inhibition rate will increase with the increasing of the concentration of OCS. When the concentration is 5mg/ml, the inhibition rate is 77%. The figure has been made with different concentrations of PPO as abscissa, the reaction rate as ordinate, when the reaction has been added OCS, the curve through the same point, has a smaller sope than the control one. This indicates that OCS is a reversible inhibitor of PPO. The double-reciprocal graph is a set of parallel lines, which indicate the OCS is the anti-competitive inhibitor of PPO.
The OCS can inhibit the browning of fresh-cut apples and improve the quality of fresh-cut apple. Although it has a lower activity of radical scavenging, it can improve the activity of POD.The results of inhibition kinetics show that the OCS is the anti-competitive inhibitor of PPO. This study indicates that the major role of OCS on fresh-cut apples is to prevent browning.
1.研究了壳寡糖对自由基的清除作用。已有相关报道证明壳寡糖对自由基具有清除作用,本研究以常用的抗氧化剂抗坏血酸和半胱氨酸为对照,对比评价壳寡糖的抗氧化性能。研究发现,壳寡糖对DPPH自由基、羟自由基、超氧阴离子都具有较好的清除作用。壳寡糖对DPPH自由基的IC50为26.06μg/ml,抗坏血酸和半胱氨酸分别是2.47μg/ml和2.43μg/ml。壳寡糖对羟自由基的IC50为0.27mg/ml,抗坏血酸和半胱氨酸分别是0.04mg/ml、0.07mg/ml。虽然抗坏血酸和半胱氨酸在低浓度条件下对超氧阴离子的清除作用优于壳寡糖,但是抗坏血酸最大清除率是50%,半胱氨酸的最大清除率是26.89%,壳寡糖的最大清除率达到了58%,说明壳寡糖对超氧阴离子的最终清除率比较高。上述结果表明壳寡糖虽然对三种自由基具有一定的清除能力,但其清除能力比抗坏血酸和半胱氨酸弱。
2.研究了壳寡糖对鲜切苹果褐变的影响。将厚度为2.5mm的鲜切苹果分别在蒸馏水、1%壳寡糖溶液、1%抗坏血酸溶液中浸泡5min,用吸水纸吸干后在10h内测定各个指标变化,以蒸馏水为阴性对照,抗坏血酸为阳性对照。研究发现,壳寡糖能够较好的抑制鲜切苹果的褐变:褐变指数在4h内明显比对照组低,与抗坏血酸组相当;多酚氧化酶活性明显比蒸馏水处理组和抗坏血酸组都要低。壳寡糖能显著提高鲜切苹果过氧化物酶(POD)含量,在10h内POD含量远远高于对照组和抗坏血酸组;壳寡糖还可以维持鲜切苹果的品质:壳寡糖处理的鲜切苹果失重率比其它两组稍低;壳寡糖对鲜切苹果的总糖含量影响不大,但可以降低可滴定酸度,这样就相应提高了糖酸比;壳寡糖还能抑制鲜切苹果表面的细菌含量,在10h内壳寡糖处理的鲜切苹果表面细菌总含量明显低于其它两组。
3.研究了壳寡糖对PPO的抑制作用。由上面的研究知道壳寡糖具有防止褐变的作用,且降低了PPO活性。本研究是在体外从反应动力学角度探讨壳寡糖对PPO的抑制作用。研究发现壳寡糖在体外对PPO有抑制作用,且随着浓度的增大抑制率增大,在5mg/ml时对PPO的抑制率为77%。以不同浓度的PPO为横坐标,以反应速率为纵坐标作图,发现加入不同浓度壳寡糖后所得到的动力学曲线经过同一点,且随着壳寡糖浓度的增大,斜率变小,说明壳寡糖是PPO的可逆性抑制剂。不同浓度壳寡糖存在条件下获得的双倒数动力曲线图得到的是一组平行线,说明壳寡糖是PPO的可逆性反竞争抑制剂。
壳寡糖可抑制鲜切苹果果肉褐变,提高贮藏期间果肉品质。尽管壳寡糖对自由基的清除作用低于抗坏血酸,但壳寡糖显著抑制了PPO的活性,且通过抑制动力学的研究发现其作用类型为反竞争性抑制。本文结果表明壳寡糖主要可作为PPO的反竞争性抑制剂抑制鲜切苹果褐变的发生。
Due to freshness and convenience, Fresh-cut fruits and vegetables are becoming more and more popular with people of all ages. But compared with the perfect ones, the quality of fresh-cut fruits and vegetables is easy to deteriorate. The research of finding safe and non-toxic natural preservation has becoming a hot topic because of people’s safety awareness. OCS, as a biosafe amino polysaccharides, also have the properties of antioxidant and antimicrobial. This study is about the antioxidant of OCS, the effect of browning of fresh-cut apples and the inbition of PPO.The details are as follow.
1. The radical scavenging activity of OCS. There have been some reports about the radical scavenging activity of OCS. This study set the ASA and cysteine as the control. The results showed that OCS has a good scavenging activity on DPPH radical, hydroxyl radical and superoxide anion. The IC50 of OCS on DPPH radical scavenging activity is 26.06ug/ml, well the ASA is 2.47ug/ml, the cysteine is 2.43ug/ml. The IC50 of OCS on hydroxyl radical scavenging is 0.27mg/ml, well the ASA is 0.04mg/ml, the cysteine is 0.07mg/ml. Although ASA and cysteine can scaveng superoxide anion at a low concentration, they did’t have a thorough scavenging ability. The maximum clearance rate of ASA is 50%, the cysteine is 26.89%. The OCS can reach 58%. In short, the OCS has a lower radical scavenging activity compared with ASA and cysteine.
2. The effect of OCS on the browning of fresh-cut apples. The fresh-cut apples which have a thickness of 2.5mm has been separately dipped in solution of distilled water, 1%ASA, 1%OCS for 5min, then, been dryed by absorbent paper. After this treatment, the fresh-cut apples were used to measure all indicators about the qulity of the apples within 10h. All the results made the ASA as positive control, and made the distilled water as negative control. This study has found that oligocatosan can inhibit the browning of fresh-cut apples. Compared with the apples treated by distilled water, the browning index of apples treated by OCS was lower within 4 hours. The OCS treatment had a lower PPO activity than the ASA and distilled water treatment. The content of POD is significantly improved within 10h compared with the other treatments. OCS can also maintain the quality of fresh-cut apples. The weight loss of OCS group is slightly lower than the other groups. The OCS has little effect on total sugar content of fresh-cut apples, but can reduce the titratable acidity, so the sugar to acid ratio has been increased. OCS can inhibit the growth of bacteria on the surface of fresh-cut apples. The content of bacteria on apples treated by OCS is significantly lower than the other groups.
3. The inhibition effect from OCS on PPO. From the study above, we know the OCS can inhibit browning and reduce the activity of PPO. The study about the inhibition from OCS on PPO is working through the method of reaction dynamics in vitro. The OCS can inhibit the activity of PPO, and the inhibition rate will increase with the increasing of the concentration of OCS. When the concentration is 5mg/ml, the inhibition rate is 77%. The figure has been made with different concentrations of PPO as abscissa, the reaction rate as ordinate, when the reaction has been added OCS, the curve through the same point, has a smaller sope than the control one. This indicates that OCS is a reversible inhibitor of PPO. The double-reciprocal graph is a set of parallel lines, which indicate the OCS is the anti-competitive inhibitor of PPO.
The OCS can inhibit the browning of fresh-cut apples and improve the quality of fresh-cut apple. Although it has a lower activity of radical scavenging, it can improve the activity of POD.The results of inhibition kinetics show that the OCS is the anti-competitive inhibitor of PPO. This study indicates that the major role of OCS on fresh-cut apples is to prevent browning.
引文
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