Journal of Zhejiang University

Journal of Zhejiang University SCIENCE B 2014 Vol.15 No.4 P.343-352

Enhanced production of thermostable laccases from a native strain of Pycnoporus sanguineus using central composite design*

Author(s): Leticia I. Ramrez-Cavazos¹,Charles Junghanns²,Rakesh Nair³,Diana L. Crdenas-Chvez¹,Carlos Hernndez-Luna⁴,Spiros N. Agathos³,Roberto Parra¹
Affiliation(s): 1. 1. Centro del Agua para Amrica Latina y el Caribe, Tecnolgico de Monterrey, Campus Monterrey, NL 64849, Mexico more
Corresponding email(s): r.parra@itesm.mx
Key Words: Enhanced laccase production, Central composite design, Screening media, Inducers, Tomato juice medium, Soybean oil, Copper sulfate

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Leticia I. Ramírez-Cavazos, Charles Junghanns, Rakesh Nair, Diana L. Cárdenas-Chávez, Carlos Hernández-Luna, Spiros N. Agathos, Roberto Parra. Enhanced production of thermostable laccases from a native strain of Pycnoporus sanguineus using central composite design[J]. Journal of Zhejiang University Science B, 2014, 15(4): 343-352.

@article{title="Enhanced production of thermostable laccases from a native strain of Pycnoporus sanguineus using central composite design",
author="Leticia I. Ramírez-Cavazos, Charles Junghanns, Rakesh Nair, Diana L. Cárdenas-Chávez, Carlos Hernández-Luna, Spiros N. Agathos, Roberto Parra",
journal="Journal of Zhejiang University Science B",
volume="15",
number="4",
pages="343-352",
year="2014",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1300246"
}

%0 Journal Article
%T Enhanced production of thermostable laccases from a native strain of Pycnoporus sanguineus using central composite design
%A Leticia I. Ramírez-Cavazos
%A Charles Junghanns
%A Rakesh Nair
%A Diana L. Cárdenas-Chávez
%A Carlos Hernández-Luna
%A Spiros N. Agathos
%A Roberto Parra
%J Journal of Zhejiang University SCIENCE B
%V 15
%N 4
%P 343-352
%@ 1673-1581
%D 2014
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1300246

TY - JOUR
T1 - Enhanced production of thermostable laccases from a native strain of Pycnoporus sanguineus using central composite design
A1 - Leticia I. Ramírez-Cavazos
A1 - Charles Junghanns
A1 - Rakesh Nair
A1 - Diana L. Cárdenas-Chávez
A1 - Carlos Hernández-Luna
A1 - Spiros N. Agathos
A1 - Roberto Parra
J0 - Journal of Zhejiang University Science B
VL - 15
IS - 4
SP - 343
EP - 352
%@ 1673-1581
Y1 - 2014
PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B1300246

Abstract
Chinese Summary
Academic Network
Reviewer Comment

Abstract: The production of thermostable laccases from a native strain of the white-rot fungus Pycnoporus sanguineus isolated in Mexico was enhanced by testing different media and a combination of inducers including copper sulfate (CuSO₄). The best conditions obtained from screening experiments in shaken flasks using tomato juice, CuSO₄, and soybean oil were integrated in an experimental design. Enhanced levels of tomato juice as the medium, CuSO₄ and soybean oil as inducers (36.8% (v/v), 3 mmol/L, and 1% (v/v), respectively) were determined for 10 L stirred tank bioreactor runs. This combination resulted in laccase titer of 143 000 IU/L (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), pH 3.0), which represents the highest activity so far reported for P. sanguineus in a 10-L fermentor. Other interesting media resulting from the screening included glucose-bactopeptone which increased laccase activity up to 20 000 IU/L, whereas the inducers Acid Blue 62 and Reactive Blue 19 enhanced enzyme production in this medium 10 times. Based on a partial characterization, the laccases of this strain are especially promising in terms of thermostability (half-life of 6.1 h at 60 °C) and activity titers.

中心组合设计方法提高血红密孔菌（Pycnoporus sanguineus）耐热漆酶的产量

研究目的：优化获得血红密孔菌（P. sanguineus）的最佳培养基组成，提高耐热漆酶的产量。
创新要点：获得了目前文献报道的最高水平的漆酶活力。
研究方法：通过单因素试验研究了不同培养基（番茄汁、麦麸、麦芽提取物和葡萄糖细菌蛋白胨培养基）和不同组合诱导剂（大豆油、阿魏酸、没食子酸、二甲基苯胺、酸性蓝62和活性蓝19分别与硫酸铜组合诱导剂）对P. sanguineus产耐热漆酶的影响。在此基础上采用中心组合试验设计，进一步研究了番茄汁培养基结合硫酸铜和大豆油组合诱导剂对P. sanguineus产耐热漆酶的影响。利用SAS10.0和响应面分析方法对试验结果进行了统计分析和建立回归模型。
重要结论：通过中心组合设计优化得出P. sanguineus产耐热漆酶的最优培养基条件：以36.8%番茄汁为培养基，以3 mmol/L硫酸铜和1%大豆油作为组合诱导剂。该条件下在10 L搅拌槽生物反应器中漆酶活力达到了143000 IU/L（2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸)为底物，pH值为3.0）。

关键词：漆酶产量；中心组合设计；培养基筛选；诱导剂；番茄汁；大豆油；硫酸铜