بررسی فعالیت و پایداری آنزیم لاکاز آزاد تثبیت شده در حضور کانی‌های مونتموریلونیت و زئولیت

نوع مقاله: مقاله پژوهشی

نویسندگان

1 دانشجوی دکتری گروه علوم خاک، دانشکده کشاورزی، دانشگاه فردوسی مشهد

2 استاد گروه علوم خاک، دانشکده کشاورزی، دانشگاه فردوسی مشهد

3 دانشیار گروه علوم خاک، دانشکده کشاورزی، دانشگاه فردوسی مشهد

4 استادیار گروه علوم خاک، دانشکده کشاورزی، دانشگاه فردوسی مشهد

چکیده

آنزیم لاکاز از گروه مهمترین آنزیم­هایی است که در سال­های گذشته به سبب توانایی آن در اکسید کردن ترکیب‌های گوناگون و آلاینده­های محیطی پایدار، نگاه بسیاری را در زمینه زیست بهسازی به سوی خود جلب کرده است. با نگاه به پایداری کم و هزینه­های ساخت و فراوری بالای آنزیم­ها، بی‌جنب­سازی آنها بر رویه نگهدارنده­ها می­تواند راهی شایسته در راستای افزایش پایداری کارکرد آنزیم باشد. برای بررسی برهم­کنش لاکاز گرفته شده از قارچ ترامتس ورسیکالر (Trametes versicolor) با کانی­های مونتموریلونیت و زئولیت سه آزمایش جداگانه در قالب طرح کاملاً تصادفی با دو تکرار در شرایط آزمایشگاهی انجام شد. آزمایش­ها به ترتیب شامل چهار سطح pH (5، 6، 7 و 8)، نه سطح دمایی (4 و80-10 درجه سانتیگراد) و هفت سطح زمان انکوباسیون (صفر، 1، 2، 5، 10، 20 و 30 روز) بودند. بر پایه آزمایش نخست، بالاترین اندازه جذب آنزیم بر رویه­های کانی­های بررسی شده در 5=pH دیده شد و با افزایش pH، جذب آنزیمی کاهش یافت. در آزمایش دوم، بالاترین اندازه فعالیت نسبی برای لاکاز آزاد در دمای 20 درجه سانتیگراد، برای لاکاز بی‌جنب­شده بر مونتموریلونیت در دماهای 80 و 4 درجه­سانتیگراد و برای لاکاز بی‌جنبش­شده بر زئولیت در دماهای 4 و 70 درجه­سانتیگراد بود. لاکاز بی جنبش­شده پایداری خوبی را در برابر دماهای پایین و بالا نشان داد. در آزمایش سوم، بالاترین اندازه فعالیت نسبی (100 درصد) برای لاکاز بی‌جنبش­شده بر مونتموریلونیت و زئولیت در زمان انکوباسیون 20 روز و برای لاکاز آزاد در زمان 5 روز دیده شد. برپایه این پژوهش، بی‌جنبش‌سازی پیامد شایسته‌ای را بر پایداری فعالیت آنزیم لاکاز نشان داد.

کلیدواژه‌ها


عنوان مقاله [English]

Studying the activity and stability of Free and immobilized Laccase enzyme (Trametes versicolor) on Montmorillonite and zeolite minerals

نویسندگان [English]

  • H. Rahmani 1
  • A. Lakzian 2
  • A. R. Karimi 3
  • A. Halajnia 4
1 PhD student, Department. of Soil Sciences, Faculty of Agriculture, Ferdowsi University of Mashhad
2 Professor, Department. of Soil Sciences, Faculty of Agriculture, Ferdowsi University of Mashhad
3 Associate professor, Department. of Soil Sciences, Faculty of Agriculture, Ferdowsi University of Mashhad
4 Assistant professor, Department. of Soil Sciences, Faculty of Agriculture, Ferdowsi University of Mashhad
چکیده [English]

Laccase is among the most important enzymes that has attracted much attention in the field of bioremediation in recent years, due to its ability to oxidize various compounds and persistent environmental pollutants. Since free from of Laccase has low operational stability with high cost of production, there are some limitations on the commercial applications in industrial and environmental biotechnology. In this regard, immobilization of such enzymes on absorbent surfaces can be a good way to increase stability and operational life of the enzyme. In order to examine the interactions between Laccase (Trametes versicolor) with Montmorillonite and zeolite minerals, three different experiments were done separately in a completely randomized design with two replications in vitro conditions. Experiments were included four levels of pH (5, 6, 7 and 8),  nine levels of temperature (4 °C and 10-80 °C) and seven levels of incubation time (0, 1, 2, 5, 10, 20 and 30 days), respectively. Based on the results, the highest enzyme adsorption on the surface of studied minerals was observed in pH=5 and enzyme absorption decreased with increasing pH. Furthermore, the highest relative activities were obtained for free Laccase at 20 °C, for immobilized Laccase on Montmorillonite at 80 and 4 °C and for immobilized Laccase on zeolite at 4 and 70 °C. Immobilized Laccase showed a remarkable stability to low and high temperatures. Maximum relative activity (100%) for immobilized Laccase on Montmorillonite and zeolite was observed in 20th day of incubation time and for free Laccase in 5th day of incubation time. Indeed, immobilization indicated a proper effect on Laccase enzyme activity retention. 

کلیدواژه‌ها [English]

  • Activity
  • Immobilization
  • laccase
  • Montmorillonite
  • Stability
  • Zeolite
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