Acyl-ACP thioesterases from macadamia (Macadamia tetraphylla) nuts: cloning, characterization and their impact on oil composition.

The mechanisms by which macadamia nuts accumulate the unusual palmitoleic and asclepic acyl moieties, which constitute up to 20% of the fatty acids in some varieties, are still unknown. Acyl-acyl carrier protein (ACP) thioesterases (EC 3.1.2.14) are intraplastidial enzymes that terminate the synthesis of fatty acids in plants and that facilitate the export of the acyl moieties to the endoplasmic reticulum where they can be used in the production of glycerolipids. Here, we have investigated the possible role of acyl-ACP thioesterase activity in the composition of macadamia kernel oil. Accordingly, two acyl-ACP thioesterases were cloned from developing macadamia kernels, one of the FatA type and the other of the FatB type. These enzymes were heterologously expressed in Escherichia coli, and the recombinant thioesterases were purified, characterized kinetically and assayed with a variety of substrates, demonstrating the high specificity of macadamia FatA towards 16:1-ACP. Acyl-ACP thioesterase activity was also characterized in crude extracts from two different varieties of macadamia, Cate and Beaumont, which accumulate different amounts of n-7 fatty acids. The impact of acyl-ACP thioesterase activities on the oil composition of these kernels is discussed in the light of these results.

Ultrastructure of Macadamia (Proteaceae) embryos: implications for their breakage properties.

BACKGROUND AND AIMS: Macadamia integrifolia, M. tetraphylla and their hybrids are cultivated for their edible kernels. Whole kernels, i.e. intact mature embryos with cotyledons fused together, are highly valued and breakage of embryos into halves results in loss of value for the commercial macadamia industry. The morphology and ultrastructure of the mature macadamia embryo, with particular emphasis on the break zone between cotyledons, were investigated. Differences in breakage between different macadamia cultivars were also examined. METHODS: Manual cracking was used to compare breakage in five cultivars and the ultrastructure of the break zone between the cotyledons was examined using light and transmission electron microscopy. KEY RESULTS: Breakage of macadamia embryos was strongly dependent on genotype of the female parent, with cultivars 'HAES 344' and 'HAES 741' much more likely to break than 'HV A16' and 'HAES 835'. Cotyledons were surrounded by a layer of cuticle resulting in a double cuticle in the break zone between the cotyledons. Three major differences have been found in the ultrastructure of the double cuticle between cultivars: a thicker cuticle in the low-whole cultivar; convolutions in the cuticle of a low-whole cultivar, and the presence of more electron-dense objects in the high-whole cultivar. CONCLUSIONS: Breakage of macadamia embryos depends on the cultivar, with clear ultrastructural differences in the break zone between cultivars. To ensure commercial benefits, macadamia breeding programs should identify germplasm with structural characteristics that ensure high percentages of whole kernel.