Molecular Characterization of a Recombinant Zea mays Phenylalanine Ammonia-Lyase (ZmPAL2) and Its Application in trans-Cinnamic Acid Production from l-Phenylalanine

Zang, Ying; Jiang, Ting; Cong, Ying; Zheng, Zhaojuan; Ouyang, Jia

doi:10.1007/s12010-015-1620-4

Cited by 22 publications

(17 citation statements)

References 44 publications

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“…Phenylalanine is the precursor in the complex phenylpropanoid pathway ( Mitchell and Walters, 1995 ; Kato et al, 2000 ; Zang et al, 2015 ). In the present study, PAL, together with CAD and POD genes, were collectively up-regulated at the early stage of Ganoderma infection (1, 2, and 3 wpi).…”

Section: Discussionmentioning

confidence: 99%

The Phenylpropanoid Pathway and Lignin in Defense against Ganoderma boninense Colonized Root Tissues in Oil Palm (Elaeis guineensis Jacq.)

et al. 2017

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Basal stem rot, caused by the basidiomycete fungus, Ganoderma boninense, is an economically devastating disease in Malaysia. Our study investigated the changes in lignin content and composition along with activity and expression of the phenylpropanoid pathway enzymes and genes in oil palm root tissues during G. boninense infection. We sampled control (non-inoculated) and infected (inoculated) seedlings at seven time points [1, 2, 3, 4, 8, and 12 weeks post-inoculation (wpi)] in a randomized design. The expression profiles of phenylalanine ammonia lyase (PAL), cinnamyl alcohol dehydrogenase (CAD), and peroxidase (POD) genes were monitored at 1, 2, and 3 wpi using real-time quantitative polymerase chain reaction. Seedlings at 4, 8, and 12 wpi were screened for lignin content, lignin composition, enzyme activities (PAL, CAD, and POD), growth (weight and height), and disease severity (DS). Gene expression analysis demonstrated up-regulation of PAL, CAD, and POD genes in the infected seedlings, relative to the control seedlings at 1, 2, and 3 wpi. At 2 and 3 wpi, CAD showed highest transcript levels compared to PAL and POD. DS increased progressively throughout sampling, with 5, 34, and 69% at 4, 8, and 12 wpi, respectively. Fresh weight and height of the infected seedlings were significantly lower compared to the control seedlings at 8 and 12 wpi. Lignin content of the infected seedlings at 4 wpi was significantly higher than the control seedlings, remained elicited with no change at 8 wpi, and then collapsed with a significant reduction at 12 wpi. The nitrobenzene oxidation products of oil palm root lignin yielded both syringyl and guaiacyl monomers. Accumulation of lignin in the infected seedlings was in parallel to increased syringyl monomers, at 4 and 8 wpi. The activities of PAL and CAD enzymes in the infected seedlings at DS = 5–34% were significantly higher than the control seedlings and thereafter collapsed at DS = 69%.

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Section: Discussionmentioning

confidence: 99%

The Phenylpropanoid Pathway and Lignin in Defense against Ganoderma boninense Colonized Root Tissues in Oil Palm (Elaeis guineensis Jacq.)

et al. 2017

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“…Only two maize PAL genes (originally named ZmPAL1 and ZmPAL2 ) have been cloned. Both were confirmed to encode proteins with PAL and tyrosine ammonia‐lyase (TAL) activities that effectively catalysed conversion of l ‐phenylalanine to trans ‐cinnamic acid (Rosler et al ., ; Zang et al ., ). It was shown that the expression of four ZmPAL genes (originally named ZmPAL1 , ZmPAL2 , ZmPAL4 and ZmPAL5 ) was strongly induced after nematode infection, while two others ( ZmPAL3 and ZmPAL6 ) were unresponsive (Starr et al ., ).…”

Section: Introductionmentioning

confidence: 97%

Maize phenylalanine ammonia‐lyases contribute to resistance to Sugarcane mosaic virus infection, most likely through positive regulation of salicylic acid accumulation

Wen

Jiang

et al. 2019

Molecular Plant Pathology

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Summary Sugarcane mosaic virus (SCMV) is a pathogen of worldwide importance that causes dwarf mosaic disease on maize (Zea mays). Until now, few maize genes/proteins have been shown to be involved in resistance to SCMV. In this study, we characterized the role of maize phenylalanine ammonia‐lyases (ZmPALs) in accumulation of the defence signal salicylic acid (SA) and in resistance to virus infection. SCMV infection significantly increased SA accumulation and expression of SA‐responsive pathogenesis‐related protein genes (PRs). Interestingly, exogenous SA treatment decreased SCMV accumulation and enhanced resistance. Both reverse transcription‐coupled quantitative PCR and RNA‐Seq data confirmed that expression levels of at least four ZmPAL genes were significantly up‐regulated upon SCMV infection. Knockdown of ZmPAL expression led to enhanced SCMV infection symptom severity and virus multiplication, and simultaneously resulted in decreased SA accumulation and PR gene expression. Intriguingly, application of exogenous SA to SCMV‐infected ZmPAL‐silenced maize plants decreased SCMV accumulation, showing that ZmPALs are required for SA‐mediated resistance to SCMV infection. In addition, lignin measurements and metabolomic analysis showed that ZmPALs are also involved in SCMV‐induced lignin accumulation and synthesis of other secondary metabolites via the phenylpropanoid pathway. In summary, our results indicate that ZmPALs are required for SA accumulation in maize and are involved in resistance to virus infection by limiting virus accumulation and moderating symptom severity.

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“…Our previous research showed that the recombinant PAL from Zea mays (ZmPAL) was completely soluble and exhibited a satisfactory PAL activity (1.78 U mg −1 crude enzyme) in E. coli . In this study, a bioconversion process of l ‐phe to t ‐CA by using engineered whole‐cell E. coli expressing ZmPAL as the biocatalyst was investigated, and the conditions for t ‐CA production were optimized.…”

Section: Resultsmentioning

confidence: 99%

“…The reported ZmPAL2 lost c . 40% of its activity when 1.40 mmol L –1 t ‐CA was added to the reaction system . Hence, it could be concluded that the existence of t ‐CA would inhibit enzyme activity and decrease the efficiency of t ‐CA production.…”

Section: Discussionmentioning

confidence: 98%

Development of a high‐efficiencytrans‐cinnamic acid bioproduction method by pH‐controlled separation technology

Zang

Xia

Zheng

et al. 2019

J of Chemical Tech & Biotech

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BACKGROUND: Trans-cinnamic acid (t-CA) and its derivatives have been shown to be antioxidant and antibacterial, and can be used in food additives and flavors. However, a traditional t-CA production via chemical synthesis with a condensation reaction may not be suitable for its application in the pharmaceutical and food fields as it is not a 'green' route. This study investigated the bioconversion of L-phenylalanine (L-phe) to t-CA using engineered whole-cell Escherichia coli BL21(DE3) expressing phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) as a biocatalyst. To prevent product inhibition, an in situ reaction and separation of t-CA was performed using a pH-controlled biotransformation strategy. RESULTS:A coupling reaction and separation system based on pH adjustment was proposed for improving the bioproduction of t-CA. Using this process, 39.61 g L −1 t-CA was bioproduced from 40 g L −1 of L-phe in 8 h and a high purity (>98%) of t-CA product was achieved directly without additional purification. CONCLUSION:The bioprocess developed in this study is a highly efficient method for the bioproduction of t-CA and, therefore, offers a promising alternative route for t-CA production. Separation of t-CA from the reaction mixture by adjusting the pHThe reaction mixture was obtained after a 2 h bioconversion using 20 g L −1 of L-phe and 8.52 g L −1 DCW of cells at pH 9.0 and 55 ∘ C. After cells were removed by centrifugation, the supernatants were adjusted to different pH values by using HCl, and were allowed to stand at room temperature for 20 min, and then the acidified supernatants were centrifuged again for separating the t-CA precipitate. The concentrations of L-phe and t-CA in the acidified supernatants at different pH values were analyzed by HPLC.

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Molecular Characterization of a Recombinant Zea mays Phenylalanine Ammonia-Lyase (ZmPAL2) and Its Application in trans-Cinnamic Acid Production from l-Phenylalanine

Cited by 22 publications

References 44 publications

The Phenylpropanoid Pathway and Lignin in Defense against Ganoderma boninense Colonized Root Tissues in Oil Palm (Elaeis guineensis Jacq.)

The Phenylpropanoid Pathway and Lignin in Defense against Ganoderma boninense Colonized Root Tissues in Oil Palm (Elaeis guineensis Jacq.)

Maize phenylalanine ammonia‐lyases contribute to resistance to Sugarcane mosaic virus infection, most likely through positive regulation of salicylic acid accumulation

Development of a high‐efficiencytrans‐cinnamic acid bioproduction method by pH‐controlled separation technology

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