AbstractStarch‐branching enzymes (SBEs) are one of the four major enzyme classes involved in starch biosynthesis in plants and algae, and their activities play a crucial role in determining the structure and physical properties of starch granules. SBEs generate α‐1,6‐branch linkages in α‐glucans through cleavage of internal α‐1,4 bonds and transfer of the released reducing ends to C‐6 hydroxyls. Starch biosynthesis in plants and algae requires multiple isoforms of SBEs and is distinct from glycogen biosynthesis in both prokaryotes and eukaryotes which uses a single branching enzyme (BE) isoform. One of the unique characteristics of starch structure is the grouping of α‐1,6‐branch points in clusters within amylopectin. This is a feature of SBEs and their interplay with other starch biosynthetic enzymes, thus facilitating formation of the compact water‐insoluble semicrystalline starch granule. In this respect, the activity of SBE isoforms is pivotal in starch granule assembly. SBEs are structurally related to the α‐amylase superfamily of enzymes, sharing three domains of secondary structure with prokaryotic Bes: the central (β/α)8‐barrel catalytic domain, an NH2‐terminal domain involved in determining the size of α‐glucan chain transferred, and the C‐terminal domain responsible for catalytic capacity and substrate preference. In addition, SBEs have conserved plant‐specific domains, including phosphorylation sites which are thought to be involved in regulating starch metabolism. SBEs form heteromeric protein complexes with other SBE isoforms as well as other enzymes involved in starch synthesis, and assembly of these protein complexes is regulated by protein phosphorylation. Phosphorylated SBEIIb is found in multienzyme complexes with isoforms of glucan‐elongating starch synthases, and these protein complexes are implicated in amylopectin cluster formation. This review presents a comparative overview of plant SBEs and includes a review of their properties, structural and functional characteristics, and recent developments on their post‐translational regulation. © 2014 IUBMB Life, 66(8):546–558, 2014