01923nas a2200265 4500008004100000022001400041245018700055210006900242260000800311520091700319653001801236653002001254653002501274653003301299653001801332653003001350100001901380700002201399700002201421700002001443700002001463700002201483700002601505856012601531 2019 eng d a0739-110200aNMR structure and dynamics of inhibitory repeat domain variant 12, a plant protease inhibitor from Capsicum annuum, and its structural relationship to other plant protease inhibitors0 aNMR structure and dynamics of inhibitory repeat domain variant 1 cAPR3 a

Although several plant protease inhibitors have been structurally characterized using X-ray crystallography, very few have been studied using NMR techniques. Here, we report an NMR study of the solution structure and dynamics of an inhibitory repeat domain (IRD) variant 12 from the wound-inducible Pin-II type proteinase inhibitor from Capsicum annuum. IRD variant 12 (IRD12) showed strong anti-metabolic activity against the Lepidopteran insect pest, Helicoverpa armigera. The NMR-derived three-dimensional structure of IRD12 reveals a three-stranded anti-parallel beta-sheet rigidly held together by four disulfide bridges and shows structural homology with known IRDs. It is interesting to note that the IRD12 structure containing similar to 75% unstructured part still shows substantial amount of rigidity of N-H bond vectors with respect to its molecular motion. Communicated by Ramaswamy H. Sarma

10aBt transgenic10aCapsicum annuum10aHelicoverpa armigera10ainhibitory repeating domains10aNMR structure10aProtease inhibitors (PIs)1 aGartia, Janeka1 aAnangi, Raveendra1 aJoshi, Rakesh, S.1 aGiri, Ashok, P.1 aKing, Glenn, F.1 aBarnwal, Ravi, P.1 aChary, Kandala, V. R. uhttp://library.ncl.res.in/content/nmr-structure-and-dynamics-inhibitory-repeat-domain-variant-12-plant-protease-inhibitor02665nas a2200241 4500008004100000022001400041245009700055210006900152260000800221300001100229490000800240520185700248653002502105653002002130653002002150653002602170653001602196100002302212700002802235700002002263700001902283856012102302 2025 eng d a0960-741200aNavigating the challenges of engineering composite specialized metabolite pathways in plants0 aNavigating the challenges of engineering composite specialized m cMAR ae701000 v1213 a

Plants are a valuable source of diverse specialized metabolites with numerous applications. However, these compounds are often produced in limited quantities, particularly under unfavorable ecological conditions. To achieve sufficient levels of target metabolites, alternative strategies such as pathway engineering in heterologous systems like microbes (e.g., bacteria and fungi) or cell-free systems can be employed. Another approach is plant engineering, which aims to either enhance the native production in the original plant or reconstruct the target pathway in a model plant system. Although increasing metabolite production in the native plant is a promising strategy, these source plants are often exotic and pose significant challenges for genetic manipulation. Effective pathway engineering requires comprehensive prior knowledge of the genes and enzymes involved, as well as the precursor, intermediate, branching, and final metabolites. Thus, a thorough elucidation of the biosynthetic pathway is closely linked to successful metabolic engineering in host or model systems. In this review, we highlight recent advances in strategies for biosynthetic pathway elucidation and metabolic engineering. We focus on efforts to engineer complex, multi-step pathways that require the expression of at least eight genes for transient and three genes for stable transformation. Reports on the engineering of complex pathways in stably transformed plants remain relatively scarce. We discuss the major hurdles in pathway elucidation and strategies for overcoming them, followed by an overview of achievements, challenges, and solutions in pathway reconstitution through metabolic engineering. Recent advances including computer-based predictions offer valuable platforms for the sustainable production of specialized metabolites in plants.

10aBiosynthetic pathway10aEnzyme activity10aGene expression10aMetabolic engineering10aMetabolites1 aGharat, Sachin, A.1 aTamhane, Vaijayanti, A.1 aGiri, Ashok, P.1 aAharoni, Asaph uhttp://library.ncl.res.in/content/navigating-challenges-engineering-composite-specialized-metabolite-pathways-plants02355nas a2200265 4500008004100000022001400041245014500055210007000200260000800270300001100278490000800289520139600297653002901693653002701722653002201749653002401771653002601795100002701821700002301848700002001871700002201891700002001913700002201933856013401955 2025 eng d a0141-813000aNovel bifunctional inhibitor of protease and α-amylase from Clitorea ternatea restricts the growth and development in Spodoptera frugiperda0 aNovel bifunctional inhibitor of protease and αamylase from Clito cMAY a1411800 v3053 a

An inhibitor molecule capable of inhibiting a wide range of digestive enzymes without affecting endogenous enzymes is always desirable. We report characterization of CteTAI (M.W. 14 kDa), a bifunctional inhibitor (BFI) protein from the seeds of Clitoria ternatea capable of inhibiting trypsin and alpha-amylase. It retains trypsin inhibition activity up to 60 degrees C and alpha-amylase inhibition up to 40 degrees C. Trypsin inhibition is stable across pH 1-12, while alpha-amylase inhibition is stable between pH 3-7. CteTAI is a noncompetitive inhibitor of trypsin and an uncompetitive inhibitor of alpha-amylase. It selectively inhibits proteases and alpha-amylases from various sources, without affecting alpha-amylase from human saliva and Bacillus spp. Proteomic analysis identified CteTAI as a bifunctional inhibitor exhibiting 41 % similarity to a bifunctional inhibitor from Sesbania bispinosa. Feeding Spodoptera frugiperda larvae with CteTAI-infused diet impaired energy metabolism, resulting in undernourished larvae and malformed adults incapable of flight and mating. Key nutritional indices (RGR, RCR, %ECI, %FDI) were severely reduced, indicating that CteTAI disrupts growth and development by inhibiting multiple protease and alpha-amylase isoforms. Biochemical characterization of newly identified CteTAI suggests its potential application in crop protection.

10aalpha-Amylase inhibitors10aBifunctional inhibitor10aClitoria ternatea10aProtease inhibitors10aSpodoptera frugiperda1 aDongare, Pratiksha, M.1 aMadage, Varsha, A.1 aDeshpande, Neha1 aJoshi, Rakesh, S.1 aGiri, Ashok, P.1 aPawar, Pankaj, K. uhttp://library.ncl.res.in/content/novel-bifunctional-inhibitor-protease-and-%CE%B1-amylase-clitorea-ternatea-restricts-growth-and