Preparation of deallergenized proteins and permuteins

An isolated deallergenized acyl lipid hydrolase protein comprising SEQ ID NO:2 modified by one or more of the following changes or SEQ ID NO:7 modified by one or more of the following changes:

• the tyrosine corresponding to position 106 of SEQ ID NO:2 or position 87 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the isoleucine corresponding to position 113 of SEQ ID NO:2 or position 94 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 129 of SEQ ID NO:2 or position 110 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the lysine corresponding to position 137 of SEQ ID NO:2 or position 118 of SEQ ID NO:7 is replaced with alanine;
• the serine corresponding to position 184 of SEQ ID NO:2 or position 165 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 185 of SEQ ID NO:2 or position 166 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the alanine corresponding to position 188 of SEQ ID NO:2 or position 169 of SEQ ID NO:7 is replaced with serine;
• the threonine corresponding to position 192 of SEQ ID NO:2 or position 173 of SEQ ID NO:7 is replaced with alanine or proline;
• the tyrosine corresponding to position 193 of SEQ ID NO:2 or position 174 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the asparagine at position 202 of SEQ ID NO:2 or position 183 of SEQ ID NO:7 is replaced with glutamine;
• the lysine corresponding to position 268 of SEQ ID NO:2 or position 249 of SEQ ID NO:7 is replaced with alanine or glutamic acid;
• the threonine corresponding to position 269 of SEQ ID NO:2 or position 250 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 270 of SEQ ID NO:2 or position 251 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the lysine corresponding to position 273 of SEQ ID NO:2 or position 254 of SEQ ID NO:7 is replaced with alanine;
• the lysine corresponding to position 313 of SEQ ID NO:2 or position 294 of SEQ ID NO:7 is replaced with glutamic acid;
• the asparagine corresponding to position 314 of SEQ ID NO:2 or position 295 of SEQ ID NO:7 is replaced with alanine;
• the asparagine corresponding to position 315 of SEQ ID NO:2 or position 296 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 316 of SEQ ID NO:2 or position 297 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the tyrosine corresponding to position 362 of SEQ ID NO:2 or position 343 of SEQ ID NO:7 is replaced with phenylalanine;
• the lysine corresponding to position 367 of SEQ ID NO:2 or position 348 of SEQ ID NO:7 is replaced with alanine;
• the arginine corresponding to position 368 of SEQ ID NO:2 or position 349 of SEQ ID NO:7 is replaced with alanine;
• the phenylalanine corresponding to position 369 of SEQ ID NO:2 or position 350 of SEQ ID NO:7 is replaced with alanine;
• the lysine corresponding to position 371 of SEQ ID NO:2 or position 352 of SEQ ID NO:7 is replaced with alanine;
• the leucine corresponding to position 372 of SEQ ID NO:2 or position 353 of SEQ ID NO:7 is replaced with alanine;
• and
• the leucine corresponding to position 373 of SEQ ID NO:2 or position 354 of SEQ ID NO:7 is replaced with alanine.

An isolated nucleic acid molecule segment comprising a structural nucleic acid sequence, the structural nucleic acid sequence encoding SEQ ID NO:2 modified in one or more of the following regions, or SEQ ID NO:7 modified in one or more of the following regions:

• positions 104-113 of SEQ ID NO:2 or positions 85-94 of SEQ ID NO:7;
• positions 128-137 of SEQ ID NO:2 or positions 109-118 of SEQ ID NO:7;
• positions 184-197 of SEQ ID NO:2 or positions 165-178 of SEQ ID NO:7;
• position 202 of SEQ ID NO:2 or position 183 of SEQ ID NO:7;
• positions 264-277 of SEQ ID NO:2 or positions 245-258 of SEQ ID NO:7;
• positions 316-325 of SEQ ID NO:2 or positions 297-306 of SEQ ID NO:7;
• and
• positions 360-377 of SEQ ID NO:2 or positions 341-358 of SEQ ID NO:7;
• wherein:
• SEQ ID NO:2 or SEQ ID NO:7 is modified by replacing one or more amino acids in the regions with alanine, glutamic acid, phenylalanine, proline, serine, or glutamine;
• and
• the modified protein displays reduced binding to anti-acyl lipid hydrolase antibodies with respect to the binding of unmodified acyl lipid hydrolase protein to the anti-acyl lipid hydrolase antibodies.

An isolated nucleic acid molecule segment comprising a structural nucleic acid sequence, the structural nucleic acid sequence encoding SEQ ID NO:2 modified by one or more of the following changes or encoding SEQ ID NO:7 modified by one or more of the following changes:

• the tyrosine corresponding to position 106 of SEQ ID NO:2 or position 87 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the isoleucine corresponding to position 113 of SEQ ID NO:2 or position 94 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 129 of SEQ ID NO:2 or position 110 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the lysine corresponding to position 137 of SEQ ID NO:2 or position 118 of SEQ ID NO:7 is replaced with alanine;
• the serine corresponding to position 184 of SEQ ID NO:2 or position 165 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 185 of SEQ ID NO:2 or position 166 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the alanine corresponding to position 188 of SEQ ID NO:2 or position 169 of SEQ ID NO:7 is replaced with serine;
• the threonine corresponding to position 192 of SEQ ID NO:2 or position 173 of SEQ ID NO:7 is replaced with alanine or proline;
• the tyrosine corresponding to position 193 of SEQ ID NO:2 or position 174 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the asparagine corresponding to position 202 of SEQ ID NO:2 or position 183 of SEQ ID NO:7 is replaced with glutamine;
• the lysine corresponding to position 268 of SEQ ID NO:2 or position 249 of SEQ ID NO:7 is replaced with alanine or glutamic acid;
• the threonine corresponding to position 269 of SEQ ID NO:2 or position 250 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 270 of SEQ ID NO:2 or position 251 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the lysine corresponding to position 273 of SEQ ID NO:2 or position 254 of SEQ ID NO:7 is replaced with alanine;
• the lysine corresponding to position 313 of SEQ ID NO:2 or position 294 of SEQ ID NO:7 is replaced with glutamic acid;
• the asparagine corresponding to position 314 of SEQ ID NO:2 or position 295 of SEQ ID NO:7 is replaced with alanine;
• the asparagine corresponding to position 315 of SEQ ID NO:2 or position 296 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 316 of SEQ ID NO:2 or position 297 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the tyrosine corresponding to position 362 of SEQ ID NO:2 or position 343 of SEQ ID NO:7 is replaced with phenylalanine;
• the lysine corresponding to position 367 of SEQ ID NO:2 or position 348 of SEQ ID NO:7 is replaced with alanine;
• the arginine corresponding to position 368 of SEQ ID NO:2 or position 349 of SEQ ID NO:7 is replaced with alanine;
• the phenylalanine corresponding to position 369 of SEQ ID NO:2 or position 350 of SEQ ID NO:7 is replaced with alanine;
• the lysine corresponding to position 371 of SEQ ID NO:2 or position 352 of SEQ ID NO:7 is replaced with alanine;
• the leucine corresponding to position 372 of SEQ ID NO:2 or position 353 of SEQ ID NO:7 is replaced with alanine;
• and
• the leucine corresponding to position 373 of SEQ ID NO:2 or position 354 of SEQ ID NO:7 is replaced with alanine.

A recombinant host cell comprising a structural nucleic acid sequence encoding SEQ ID NO:2 modified in one or more of the following regions, or SEQ ID NO:7 modified in one or more of the following regions:

• positions 104-113 of SEQ ID NO:2 or positions 85-94 of SEQ ID NO:7;
• positions 128-137 of SEQ ID NO:2 or positions 109-118 of SEQ ID NO:7;
• positions 184-197 of SEQ ID NO:2 or positions 165-178 of SEQ ID NO:7;
• position 202 of SEQ ID NO:2 or position 183 of SEQ ID NO:7;
• positions 264-277 of SEQ ID NO:2 or positions 245-258 of SEQ ID NO:7;
• positions 316-325 of SEQ ID NO:2 or positions 297-306 of SEQ ID NO:7;
• and
• positions 360-377 of SEQ ID NO:2 or positions 341-358 of SEQ ID NO:7;
• wherein:
• SEQ ID NO:2 or SEQ ID NO:7 is modified by replacing one or more amino acids in the regions with alanine, glutamic acid, phenylalanine, proline, serine, or glutamine;
• and
• the modified protein displays reduced binding to anti-acyl lipid hydrolase antibodies with respect to the binding of unmodified acyl lipid hydrolase protein to the anti-acyl lipid hydrolase antibodies.

A recombinant host cell comprising a structural nucleic acid sequence encoding SEQ ID NO:2 modified by one or more of the following changes or encoding SEQ ID NO:7 modified by one or more of the following changes:

• the tyrosine corresponding to position 106 of SEQ ID NO:2 or position 87 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the isoleucine corresponding to position 113 of SEQ ID NO:2 or position 94 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 129 of SEQ ID NO:2 or position 110 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the lysine corresponding to position 137 of SEQ ID NO:2 or position 118 of SEQ ID NO:7 is replaced with alanine;
• the serine corresponding to position 184 of SEQ ID NO:2 or position 165 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 185 of SEQ ID NO:2 or position 166 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the alanine corresponding to position 188 of SEQ ID NO:2 or position 169 of SEQ ID NO:7 is replaced with serine;
• the threonine corresponding to position 192 of SEQ ID NO:2 or position 173 of SEQ ID NO:7 is replaced with alanine or proline;
• the tyrosine corresponding to position 193 of SEQ ID NO:2 or position 174 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the asparagine corresponding to position 202 of SEQ ID NO:2 or position 183 of SEQ ID NO:7 is replaced with glutamine;
• the lysine corresponding to position 268 of SEQ ID NO:2 or position 249 of SEQ ID NO:7 is replaced with alanine or glutamic acid;
• the threonine corresponding to position 269 of SEQ ID NO:2 or position 250 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 270 of SEQ ID NO:2 or position 251 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the lysine corresponding to position 273 of SEQ ID NO:2 or position 254 of SEQ ID NO:7 is replaced with alanine;
• the lysine corresponding to position 313 of SEQ ID NO:2 or position 294 of SEQ ID NO:7 is replaced with glutamic acid;
• the asparagine corresponding to position 314 of SEQ ID NO:2 or position 295 of SEQ ID NO:7 is replaced with alanine;
• the asparagine corresponding to position 315 of SEQ ID NO:2 or position 296 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 316 of SEQ ID NO:2 or position 297 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the tyrosine corresponding to position 362 of SEQ ID NO:2 or position 343 of SEQ ID NO:7 is replaced with phenylalanine;
• the lysine corresponding to position 367 of SEQ ID NO:2 or position 348 of SEQ ID NO:7 is replaced with alanine;
• the arginine corresponding to position 368 of SEQ ID NO:2 or position 349 of SEQ ID NO:7 is replaced with alanine;
• the phenylalanine corresponding to position 369 of SEQ ID NO:2 or position 350 of SEQ ID NO:7 is replaced with alanine;
• the lysine corresponding to position 371 of SEQ ID NO:2 or position 352 of SEQ ID NO:7 is replaced with alanine;
• the leucine corresponding to position 372 of SEQ ID NO:2 or position 353 of SEQ ID NO:7 is replaced with alanine;
• and
• the leucine corresponding to position 373 of SEQ ID NO:2 or position 354 of SEQ ID NO:7 is replaced with alanine.

A recombinant plant comprising a structural nucleic acid sequence encoding SEQ ID NO:2 modified in one or more of the following regions, or SEQ ID NO:7 modified in one or more of the following regions:

• positions 104-113 of SEQ ID NO:2 or positions 85-94 of SEQ ID NO:7;
• positions 128-137 of SEQ ID NO:2 or positions 109-118 of SEQ ID NO:7;
• positions 184-197 of SEQ ID NO:2 or positions 165-178 of SEQ ID NO:7;
• position 202 of SEQ ID NO:2 or position 183 of SEQ ID NO:7;
• positions 264-277 of SEQ ID NO:2 or positions 245-258 of SEQ ID NO:7;
• positions 316-325 of SEQ ID NO:2 or positions 297-306 of SEQ ID NO:7;
• and
• positions 360-377 of SEQ ID NO:2 or positions 341-358 of SEQ ID NO:7;
• wherein:
• SEQ ID NO:2 or SEQ ID NO:7 is modified by replacing one or more amino acids in the regions with alanine, glutamic acid, phenylalanine, proline, serine, or glutamine;
• and
• the modified protein displays reduced binding to anti-acyl lipid hydrolase antibodies with respect to the binding of unmodified acyl lipid hydrolase protein to the anti-acyl lipid hydrolase antibodies.

A recombinant plant comprising a structural nucleic acid sequence encoding SEQ ID NO:2 modified by one or more of the following changes or encoding SEQ ID NO:7 modified by one or more of the following changes:

• the tyrosine corresponding to position 106 of SEQ ID NO:2 or position 87 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the isoleucine corresponding to position 113 of SEQ ID NO:2 or position 94 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 129 of SEQ ID NO:2 or position 110 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the lysine corresponding to position 137 of SEQ ID NO:2 or position 118 of SEQ ID NO:7 is replaced with alanine;
• the serine corresponding to position 184 of SEQ ID NO:2 or position 165 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 185 of SEQ ID NO:2 or position 166 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the alanine corresponding to position 188 of SEQ ID NO:2 or position 169 of SEQ ID NO:7 is replaced with serine;
• the threonine corresponding to position 192 of SEQ ID NO:2 or position 173 of SEQ ID NO:7 is replaced with alanine or proline;
• the tyrosine corresponding to position 193 of SEQ ID NO:2 or position 174 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the asparagine corresponding to position 202 of SEQ ID NO:2 or position 183 of SEQ ID NO:7 is replaced with glutamine;
• the lysine corresponding to position 268 of SEQ ID NO:2 or position 249 of SEQ ID NO:7 is replaced with alanine or glutamic acid;
• the threonine corresponding to position 269 of SEQ ID NO:2 or position 250 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 270 of SEQ ID NO:2 or position 251 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the lysine corresponding to position 273 of SEQ ID NO:2 or position 254 of SEQ ID NO:7 is replaced with alanine;
• the lysine corresponding to position 313 of SEQ ID NO:2 or position 294 of SEQ ID NO:7 is replaced with glutamic acid;
• the asparagine corresponding to position 314 of SEQ ID NO:2 or position 295 of SEQ ID NO:7 is replaced with alanine;
• the asparagine corresponding to position 315 of SEQ ID NO:2 or position 296 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 316 of SEQ ID NO:2 or position 297 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the tyrosine corresponding to position 362 of SEQ ID NO:2 or position 343 of SEQ ID NO:7 is replaced with phenylalanine;
• the lysine corresponding to position 367 of SEQ ID NO:2 or position 348 of SEQ ID NO:7 is replaced with alanine;
• the arginine corresponding to position 368 of SEQ ID NO:2 or position 349 of SEQ ID NO:7 is replaced with alanine;
• the phenylalanine corresponding to position 369 of SEQ ID NO:2 or position 350 of SEQ ID NO:7 is replaced with alanine;
• the lysine corresponding to position 371 of SEQ ID NO:2 or position 352 of SEQ ID NO:7 is replaced with alanine;
• the leucine corresponding to position 372 of SEQ ID NO:2 or position 353 of SEQ ID NO:7 is replaced with alanine;
• and
• the leucine corresponding to position 373 of SEQ ID NO:2 or position 354 of SEQ ID NO:7 is replaced with alanine.

A method of preparing a recombinant plant transformed to produce a protein modified to exhibit reduced allergen eliciting properties when consumed in the diet of a human allergic to the unmodified protein, the method comprising:

• selecting a host plant cell;
• transforming the host plant cell with a recombinant vector;
• obtaining recombinant host cells;
• and
• regenerating a recombinant plant from the recombinant host plant cells;
• wherein the recombinant vector comprises a structural nucleic acid sequence encoding SEQ ID NO:2 modified in one or more of the following regions, or SEQ ID NO:7 modified in one or more of the following regions:
• positions 104-113 of SEQ ID NO:2 or positions 85-94 of SEQ ID NO:7;
• positions 128-137 of SEQ ID NO:2 or positions 109-118 of SEQ ID NO:7;
• positions 184-197 of SEQ ID NO:2 or positions 165-178 of SEQ ID NO:7;
• position 202 of SEQ ID NO:2 or position 183 of SEQ ID NO:7;
• positions 264-277 of SEQ ID NO:2 or positions 245-258 of SEQ ID NO:7;
• positions 316-325 of SEQ ID NO:2 or positions 297-306 of SEQ ID NO:7;
• and
• positions 360-377 of SEQ ID NO:2 or positions 341-358 of SEQ ID NO:7;
• wherein:
• SEQ ID NO:2 or SEQ ID NO:7 is modified by replacing one or more amino acids in the regions with alanine, glutamic acid, phenylalanine, proline, serine, or glutamine;
• and
• the modified protein displays reduced binding to anti-acyl lipid hydrolase antibodies with respect to the binding of unmodified acyl lipid hydrolase protein to the anti-acyl lipid hydrolase antibodies.

A method of preparing a recombinant plant transformed to produce a protein modified to exhibit reduced allergen eliciting properties when consumed in the diet of a human allergic to the unmodified protein, the method comprising:

• selecting a host plant cell;
• transforming the host plant cell with a recombinant vector;
• obtaining recombinant host cells;
• and
• regenerating a recombinant plant from the recombinant host plant cells;
• wherein the recombinant vector comprises a structural nucleic acid sequence encoding SEQ ID NO:2 modified by one or more of the following changes or encoding SEQ ID NO:7 modified by one or more of the following changes:
• the tyrosine corresponding to position 106 of SEQ ID NO:2 or position 87 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the isoleucine corresponding to position 113 of SEQ ID NO:2 or position 94 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 129 of SEQ ID NO:2 or position 110 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the lysine corresponding to position 137 of SEQ ID NO:2 or position 118 of SEQ ID NO:7 is replaced with alanine;
• the serine corresponding to position 184 of SEQ ID NO:2 or position 165 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 185 of SEQ ID NO:2 or position 166 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the alanine corresponding to position 188 of SEQ ID NO:2 or position 169 of SEQ ID NO:7 is replaced with serine;
• the threonine corresponding to position 192 of SEQ ID NO:2 or position 173 of SEQ ID NO:7 is replaced with alanine or proline;
• the tyrosine corresponding to position 193 of SEQ ID NO:2 or position 174 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the asparagine corresponding to position 202 of SEQ ID NO:2 or position 183 of SEQ ID NO:7 is replaced with glutamine;
• the lysine corresponding to position 268 of SEQ ID NO:2 or position 249 of SEQ ID NO:7 is replaced with alanine or glutamic acid;
• the threonine corresponding to position 269 of SEQ ID NO:2 or position 250 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 270 of SEQ ID NO:2 or position 251 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the lysine corresponding to position 273 of SEQ ID NO:2 or position 254 of SEQ ID NO:7 is replaced with alanine;
• the lysine corresponding to position 313 of SEQ ID NO:2 or position 294 of SEQ ID NO:7 is replaced with glutamic acid;
• the asparagine corresponding to position 314 of SEQ ID NO:2 or position 295 of SEQ ID NO:7 is replaced with alanine;
• the asparagine corresponding to position 315 of SEQ ID NO:2 or position 296 of SEQ ID NO:7 is replaced with alanine;
• the tyrosine corresponding to position 316 of SEQ ID NO:2 or position 297 of SEQ ID NO:7 is replaced with phenylalanine or alanine;
• the tyrosine corresponding to position 362 of SEQ ID NO:2 or position 343 of SEQ ID NO:7 is replaced with phenylalanine;
• the lysine corresponding to position 367 of SEQ ID NO:2 or position 348 of SEQ ID NO:7 is replaced with alanine;
• the arginine corresponding to position 368 of SEQ ID NO:2 or position 349 of SEQ ID NO:7 is replaced with alanine;
• the phenylalanine corresponding to position 369 of SEQ ID NO:2 or position 350 of SEQ ID NO:7 is replaced with alanine;
• the lysine corresponding to position 371 of SEQ ID NO:2 or position 352 of SEQ ID NO:7 is replaced with alanine;
• the leucine corresponding to position 372 of SEQ ID NO:2 or position 353 of SEQ ID NO:7 is replaced with alanine;
• and
• the leucine corresponding to position 373 of SEQ ID NO:2 or position 354 of SEQ ID NO:7 is replaced with alanine.

A method of preparing a recombinant host cell, the method comprising:

• selecting a host cell;
• transforming the host cell with a recombinant vector;
• and
• obtaining recombinant host cells;
• wherein the recombinant vector comprises a structural nucleic acid sequence encoding a protein selected from the group consisting of SEQ ID NO:247 and SEQ ID NO:260.

A method of preparing a recombinant plant, the method comprising:

• selecting a host plant cell;
• transforming the host plant cell with a recombinant vector;
• obtaining recombinant host cells;
• and
• regenerating a recombinant plant from the recombinant host plant cells;
• wherein the recombinant vector comprises a structural nucleic acid sequence encoding a protein selected from the group consisting of SEQ ID NO:274 and SEQ ID NO:260.

An isolated protein comprising:

• (amino acids 247-386 of SEQ ID NO:2)-linker-(amino acids 24-246 of SEQ ID NO:2) or
• (amino acids 269-386 of SEQ ID NO:2)-linker-(amino acids 24-268 of SEQ ID NO:2);
• the protein modified by replacing one or more amino acids in the following regions with alanine, glutamic acid, phenylalanine, proline, serine, or glutamine:
• positions 104-113 of SEQ ID NO:2;
• positions 128-137 of SEQ ID NO:2;
• positions 184-197 of SEQ ID NO:2;
• position 202 of SEQ ID NO:2;
• positions 264-277 of SEQ ID NO:2;
• positions 316-325 of SEQ ID NO:2;
• and
• positions 360-377 of SEQ ID NO:2;
• wherein the protein displays reduced binding to anti-acyl lipid hydrolase antibodies with respect to the binding of unmodified acyl lipid hydrolase protein to the anti-acyl lipid hydrolase antibodies;
• and
• wherein said linker is selected from the group consisting of SEQ ID NO:277 and SEQ ID NO:276;
• and

An isolated nucleic acid molecule segment comprising a structural nucleic acid sequence, the structural nucleic acid sequence encoding a protein selected from the group consisting of (amino acids 247-386 of SEQ ID NO:2)-linker-(amino acids 24-246 of SEQ ID NO:2);

• and
• (amino acids 269-386 of SEQ ID NO:2)-linker-(amino acids 24-268 of SEQ ID NO:2);
• the protein modified by replacing one or more amino acids in the following regions with alanine, glutamic acid, phenylalanine, proline, serine, or glutamine:
• positions 104-113 of SEQ ID NO:2;
• positions 128-137 of SEQ ID NO:2;
• positions 184-197 of SEQ ID NO:2;
• position 202 of SEQ ID NO:2;
• positions 264-277 of SEQ ID NO:2;
• positions 316-325 of SEQ ID NO:2;
• and
• positions 360-377 of SEQ ID NO:2;
• wherein the protein displays reduced binding to anti-acyl lipid hydrolase antibodies with respect to the binding of unmodified acyl lipid hydrolase protein to the anti-acyl lipid hydrolase antibodies;
• and
• wherein the linker is selected from the group consisting of SEQ ID NO:277 and SEQ ID NO:276;
• and

A recombinant vector comprising operatively linked in the 5′ to 3′ orientation:

• a promoter that directs transcription of a structural nucleic acid sequence;
• a structural nucleic acid sequence encoding a protein selected from the group consisting of (amino acids 247-386 of SEQ ID NO:2)-linker-(amino acids 24-246 of SEQ ID NO:2);
• and
• (amino acids 269-386 of SEQ ID NO:2)-linker-(amino acids 24-268 of SEQ ID NO:2);
• and
• a 3′ transcription terminator;
• the protein modified by replacing one or more amino acids in the following regions with alanine, glutamic acid, phenylalanine, proline, serine, or glutamine:
• positions 104-113 of SEQ ID NO:2;
• positions 128-137 of SEQ ID NO:2;
• positions 184-197 of SEQ ID NO:2;
• position 202 of SEQ ID NO:2;
• positions 264-277 of SEQ ID NO:2;
• positions 316-325 of SEQ ID NO:2;
• and
• positions 360-377 of SEQ ID NO:2;
• wherein said linker is selected from the group consisting of SEQ ID NO:277 and SEQ ID NO:276;
• and
• wherein the protein displays reduced binding to anti-acyl lipid hydrolase antibodies with respect to the binding of unmodified acyl lipid hydrolase protein to the anti-acyl lipid hydrolase antibodies.

A recombinant host cell comprising a structural nucleic acid sequence encoding a protein selected from the group consisting of (amino acids 247-386 of SEQ ID NO:2)-linker-(amino acids 24-246 of SEQ ID NO:2);

• and
• (amino acids 269-386 of SEQ ID NO:2)-linker-(amino acids 24-268 of SEQ ID NO:2);
• the protein modified by replacing one or more amino acids in the following regions with alanine, glutamic acid, phenylalanine, proline, serine, or glutamine:
• positions 104-113 of SEQ ID NO:2;
• positions 128-137 of SEQ ID NO:2;
• positions 184-197 of SEQ ID NO:2;
• position 202 of SEQ ID NO:2;
• positions 264-277 of SEQ ID NO:2;
• positions 316-325 of SEQ ID NO:2;
• and
• positions 360-377 of SEQ ID NO:2;
• wherein said linker is selected from the group consisting of SEQ ID NO:277 and SEQ ID NO:276;
• and
• wherein the protein displays reduced binding to anti-acyl lipid hydrolase antibodies with respect to the binding of unmodified acyl lipid hydrolase protein to the anti-acyl lipid hydrolase antibodies.

A recombinant plant comprising a structural nucleic acid sequence encoding a protein selected from the group consisting of (amino acids 247-386 of SEQ ID NO:2)-linker-(amino acids 24-246 of SEQ ID NO:2);

• and
• (amino acids 269-386 of SEQ ID NO:2)-linker-(amino acids 24-268 of SEQ ID NO:2);
• the protein modified by replacing one or more amino acids in the following regions with alanine, glutamic acid, phenylalanine, proline, serine, or glutamine:
• positions 104-113 of SEQ ID NO:2;
• positions 128-137 of SEQ ID NO:2;
• positions 184-197 of SEQ ID NO:2;
• position 202 of SEQ ID NO:2;
• positions 264-277 of SEQ ID NO:2;
• positions 316-325 of SEQ ID NO:2;
• and
• positions 360-377 of SEQ ID NO:2;
• wherein said linker is selected from the group consisting of SEQ ID NO:277 and SEQ ID NO:276;
• and
• wherein the protein displays reduced binding to anti-acyl lipid hydrolase antibodies with respect to the binding of unmodified acyl lipid hydrolase protein to the anti-acyl lipid hydrolase antibodies.

A method for decreasing allergen eliciting properties of a native protein comprising the steps of:

• a) identifying a patient exhibiting an allergic sensitivity to said native protein and obtaining serum from said patient;
• b) exposing synthetic overlapping peptides representative of said native protein to said patient serum to identify peptides exhibiting epitopes which bind IgE present within said patient serum;
• c) producing variant peptides exhibiting alanine scanning or rational scanning amino acid substitutions based on peptides from step (b), wherein said variant peptides exhibit decreased IgE binding compared to peptides from step (b), said amino acid substitutions comprising result effective substitutions;
• d) modifying the amino acid sequence of said native protein to contain one or more of said result effective substitutions;
• and
• e) isolating and purifying the modified protein comprising one or more result effective amino acid substitutions;
• wherein the modified protein comprising said one or more result effective substitutions exhibits reduced binding of IgE present within said patient serum when compared with said native protein.

A method for producing a modified acyl lipid hydrolase protein comprising:

• a) identifying a patient exhibiting an allergic sensitivity to a native form of said hydrolase protein and obtaining serum from said patient;
• b) exposing synthetic overlapping peptides representative of said native protein to said patient serum to identify peptides exhibiting epitopes which bind immunoglobulins present within said patient serum, said immunoglobulins exhibiting a binding specificity for said native protein;
• c) roducing variant peptides exhibiting alanine scanning or rational scanning amino acid substitutions based on peptides from step (b), wherein said variant peptides exhibit decreased immunoglobulin binding compared to peptides from step (b), said amino acid substitutions comprising result effective substitutions;
• d) modifying the amino acid sequence of said native protein to contain one or more of said result effective substitutions;
• and
• e) isolating and purifying the modified protein comprising one or more result effective amino acid substitutions;
• wherein the modified acyl lipid hydrolase protein comprises said one or more result effective substitutions, exhibits reduced binding of immunoglobulins present within said patient serum when compared with said native protein, exhibits lipid acyl hydrolase activity no less than that of the native protein, and exhibits insecticidal activity no less than that of the native protein.